Inhalant Formulation Containing Sulfoalkyl Ether Cyclodextrin and Corticosteroid

ABSTRACT

An inhalable formulation containing SAE-CD and corticosteroid is provided. The formulation is adapted for administration to a subject by nebulization with any known nebulizer. The formulation can be included in a kit. The formulation is administered as an aqueous solution, however, it can be stored as a dry powder, ready-to-use solution, or concentrated composition. The formulation is employed in an improved nebulization system for administering corticosteroid by inhalation. SAE-CD present in the formulation significantly enhances the chemical stability of budesonide. A method of administering the formulation by inhalation is provided. The formulation can also be administered by conventional nasal delivery apparatus.

CROSS-REFERENCE TO EARLIER FILED APPLICATIONS

The present application is a continuation-in-part of and claims thepriority of U.S. application Ser. No. 11/479,979 filed Jun. 30, 2006,which is a continuation-in-part of PCT International Application No.PCT/US05/00082 filed Dec. 31, 2004, which claims benefit of the filingdate of provisional application No. 60/533,628 filed Dec. 31, 2003, thedisclosures of which are hereby incorporated by reference in theirentirety.

FIELD OF THE INVENTION

The present invention relates to methods of and systems foradministering formulations of sulfoalkyl ether cyclodextrin andcorticosteroid, such as budesonide, by nebulization and inhalation. Theinvention also relates to methods of treating respiratory diseases ordisorders of the air passageways.

BACKGROUND OF THE INVENTION

The delivery of a drug by inhalation allows deposition of the drug indifferent sections of the respiratory tract, e.g., throat, trachea,bronchi and alveoli. Generally, the smaller the particle size, thelonger the particle will remain suspended in air and the farther downthe respiratory tract the drug can be delivered. Corticosteroids aredelivered by inhalation using nebulizers, metered dose inhalers, or drypowder inhalers. The principle advantages of nebulizers over othermethods of pulmonary installation are that patient cooperation is notrequired and the delivery of higher doses of medication is easier. Themain concerns about nebulizers, however, are their increased cost,reduced portability and the inconvenience of needing to preparemedication beforehand and the increased time requirement foradministering a treatment. A method of improving the administration ofdrugs, such as corticosteroids by nebulization would be desired.

Budesonide ((R,S)-11β, 16α, 17, 21-tetrahydroxypregna-1, 4-diene-3,20-dione cyclic 16, 17-acetal with butyraldehyde; C₂₅H₃₄O₆; Mw: 430.5)is well known. It is provided commercially as a mixture of two isomers(22R and 22S). Budesonide is an anti-inflammatory corticosteroid thatexhibits potent glucocorticoid activity. Administration of budesonide isindicated for maintenance treatment of asthma and as prophylactictherapy in children.

Commercial formulations of budesonide are sold by AstraZeneca LP(Wilmington, Del.) under the trademarks ENTOCORT™ EC, PULMICORTRESPULES®, Rhinocort Aqua®, Rhinocort® Nasal Inhaler and PulmicortTurbuhaler®, and under its generic name. PULMICORT RESPULES® suspension,which is a sterile aqueous suspension of micronized budesonide, isadministered by inhalation using a nebulizer, in particular a compressedair driven jet nebulizer that delivers from 2 to 18% of the drug masscontained in the nominal charge. The general formulation for a unit doseof the PULMICORT RESPULES is set forth in U.S. Pat. No. 6,598,603, andit is an aqueous suspension in which budesonide is suspended in anaqueous medium comprising about 0.05 to 1.0 mg of budesonide, 0.05 to0.15 mg of NaEDTA, 8.0 to 9.0 mg of NaCl, 0.15 to 0.25 mg ofpolysorbate, 0.25 to 0.30 mg of anhydrous citric acid, and 0.45 to 0.55mg of sodium citrate per one ml of water. RHINOCORT® NASAL INHALER™ is ametered-dose pressurized aerosol unit containing a suspension ofmicronized budesonide in a mixture of propellants. RHINOCORT® AQUA™ isan unscented metered-dose manual-pump spray formulation containing asuspension of micronized budesonide in an aqueous medium. Thesuspensions should not be administered with an ultrasonic nebulizer.

A wide variety of nebulizers differing in mode of operation areavailable, e.g. jet nebulizers (optionally sold with compressors),ultrasonic nebulizers, vibrating membrane, vibrating mesh nebulizers,vibrating plate nebulizers, vibrating cone nebulizers, and others. Thevibrating mesh, vibrating cone or vibrating plate nebulizers are ofparticular interest since they do not require the use of an aircompressor for delivery, have a minimal residual volume in the reservoirafter delivery of a unit dose, and can be used to deliver low volumes ofinhalable solutions. Exemplary vibrating membrane, mesh or platenebulizers are described by R. Dhand (Respiratory Care, (December 2002),47(12), p. 1406-1418). Keller et al. (ATS 99^(th) InternationalConference, Seattle, May 16^(th)-21^(st), 2003; poster 2727) disclosethe results of a study on the administration of AZTREONAM with a PARIeFlow nebulizer and report low oropharyngeal deposition, nebulizationefficiency unaffected by fill volume, a respirable fraction of 82±1.7%,constant dose administration per inhalation cycle time, and an excellentcorrelation between delivered dose, fill volume and nebulization time,and expected high lung deposition.

The desired properties of a liquid for nebulization generallyinclude: 1) reduced viscosity; 2) sterile medium; 3) reduced surfacetension; 4) stability toward the mechanism of the nebulizer; 5) moderatepH of about 4-10; 6) ability to form droplets with an MMAD of <5 μm orpreferably <3 μm; 7) absence of irritating preservatives and stabilizingagents; 8) suitable tonicity. On the one hand, suspensions possess someadvantages but on the other hand solutions possess other advantages.

Smaldone et al. (J. Aerosol Med. (1998), 11, 113-125) disclose theresults of a study on the in vitro determination of inhaled mass andparticle distribution of a budesonide suspension. They conclude that2%-18% of the nebulizer's charge of budesonide was delivered from thesuspension, meaning that budesonide delivery was incomplete resulting ina significant waste of drug. In the thirteen most efficient systems, thesuspension can be nebulized sufficiently well for lower respiratorytract delivery.

Another study further demonstrated the highly variable efficiency ofnebulization from one nebulizer to another. Barry et al. (J. AllergyClin. Immunol. (1998), 320-321) state that this variability should betaken into account when treating patients with nebulized budesonide.Berg et al. (J. Aerosol Sci. (1998), 19(7), 1101-1104) also report thehighly variable efficiency of nebulization of PULMICORT™ suspension fromone nebulizer to the next. Moreover, the mass mean aerodynamic diameter(MMAD) of the nebulized droplets is highly variable from one nebulizerto the next. In general, suspensions are less efficiently nebulized thansolutions, O'Riordan (Respiratory Care, (2002), 1305-1313). Inhaledcorticosteroids are utilized in the treatment of asthma and are ofsignificant benefit because they are delivered directly to the site ofaction, the lung. The goal of an inhaled corticosteroid is to providelocalized therapy with immediate drug activity in the lungs. Inhaledcorticosteroids are well absorbed from the lungs. In fact, it can beassumed that all of the drug available at the receptor site in the lungswill be absorbed systemically. However, it is well known that usingcurrent methods and formulations the greater part of an inhaledcorticosteroid dose is swallowed and becomes available for oralabsorption, resulting in unwanted systemic effects. For inhaledcorticosteroids, high pulmonary availability is more important than highoral bioavailability because the lung is the target organ. A productwith high pulmonary availability has greater potential to exert positiveeffects in the lung. The ideal inhaled corticosteroid formulation wouldprovide minimum oral delivery thereby reducing the likelihood ofsystemic adverse effects.

The majority of the corticosteroid dose delivered to the lung isabsorbed and available systemically. For the portion of the inhaledcorticosteroid dose delivered orally, bioavailability depends uponabsorption from the GI tract and the extent of first pass metabolism inthe liver. Since this oral component of corticosteroid drug deliverydoes not provide any beneficial therapeutic effect but can increasesystemic side effects, it is desirable for the oral bioavailability ofinhaled corticosteroid to be relatively low.

Both particle size and formulation influence the efficacy of an inhaledcorticosteroid. The formulation of a drug has a significant impact onthe delivery of that drug to the lungs, and therefore its efficacy. Mostimportant in the delivery of drug to the lung are the aerosol vehicleand the size of the particles delivered. Additionally, a reduced degreeof pulmonary deposition suggests a greater degree of oropharyngealdeposition. Due to a particular formulation employed, somecorticosteroids are more likely to be deposited in the mouth and throatand may cause local adverse effects.

While receptor distribution is the major determinant of bronchodilatorefficacy, particle size appears to be more important in determining theefficacy of an inhaled corticodsteroid. The smallest airways have aninternal diameter of 2 micrometers (mcm) or less. Thus, an inhaler withparticles having a mean aerodynamic diameter of 1 mcm should have agreater respirable fraction than an inhaler with particles that have anaverage diameter of 3.5 to 4 mcm. For patients with obstructive lungdisease, all particles should ideally be no greater than 2 to 3 mcm. Aparticle that is small (less than 5 mcm) is more likely to be inhaledinto the smaller airways of the lungs, thus improving efficacy. Incontrast, particles that are larger than 5 mcm can be deposited in themouth and throat, both reducing the proportion of particles that reachthe lungs and potentially causing local adverse effects such as oralcandidiasis and hoarseness (dysphonia). Particles having a mass medianaerodynamic diameter (MMAD) of close to 1 mcm are considered to have agreater respirable fraction per dose than those with a diameter of 3.5mcm or greater.

A further disadvantage to the nebulization of budesonide suspensions isthe need to generate very small droplets, MMAD of about <3 μm. Since thenebulized droplets are so small, then the micronized budesonide must beeven smaller or in the range of 0.5-2.0 μm and the particles should havea narrow particle size distribution. Generation of such particles isdifficult.

Even so, efforts have been made to improve the nebulization ofbudesonide suspensions with ultrasonic nebulizers by usingsubmicron-sized particles (Keller et al. in Respiratory Drug DeliveryVIII (2002), 197-206). A suspension of nanoparticles (0.1-1.0 μm) of thecorticosteroid might be used to increase the proportion of respirableparticles as compared to a coarser suspension as in the PULMICORT™suspension. No improvement over PULMICORT™ suspension (about 4.4 μmbudesonide particle size in suspension) was observed. Moreover, concernsexist regarding the use of nanosuspensions in that the small particles(<0.05 μm) may induce an allergic response in a subject. SheffieldPharmaceuticals, Inc. (St. Louis, Mo.; “The Pharmacokinetics ofNebulized Nanocrystal Budesonide Suspension in Healthy Volunteers”,Kraft, et al. in J. Clin. Pharmacol., (2004), 44:67-72) has disclosedthe preparation and evaluation of UDB (unit dose budesonide), which is asuspension-based formulation containing nanoparticles of budesonidedispersed in a liquid medium. This product is being developed by MAPPharmaceuticals, Inc. (Mountain View, Calif.). Seeman et al. (ATS99^(th) International Conference, Seattle, May 16^(th)-21^(st), 2003;poster 2727) disclose the results of a study evaluating the performanceof the PARI eFlow nebulizer with a budesonide suspension (PULMICORTRESPULES, 500 μg/ml, in a 2 ml ampoule) and report achieving a MMAD of3.6-4.2 μm and a respirable fraction of greater than 67%.

The inhalation of drug particles as opposed to dissolved drug is knownto be disadvantageous. Brain et al. (Bronchial Asthma, 2^(nd) Ed. (Ed.E. B. Weis et al., Little Brown & Co. (1985), pp. 594-603) report thatless soluble particles that deposit on the mucous blanket coveringpulmonary airways and the nasal passages are moved toward the pharynx bythe cilia. Such particles would include the larger drug particlesdeposited in the upper respiratory tract. Mucus, cells and debris comingfrom the nasal cavities and the lungs meet at the pharynx, mix withsaliva, and enter the gastrointestinal tract upon being swallowed.Reportedly, by this mechanism, particles are removed from the lungs withhalf-times of minutes to hours. Accordingly, there is little time forsolubilization of slowly dissolving drags, such as budesonide. Incontrast, particles deposited in the nonciliated compartments, such asthe alveoli, have much longer residence times. Since it is difficult togenerate very small particles of budesonide for deep lung deposition,much of the inhaled suspension would likely be found in the upper tomiddle respiratory tract. However, it is much easier to generate smalldroplets from a solution than it is from a suspension of solids. Forthese reasons, nebulization of a budesonide-containing solution shouldbe preferred over that of a suspension.

O'Riordan (Respiratory Care (2002 November), 47(11), 1305-1313) statesthat drugs can be delivered by nebulization of either solutions orsuspensions, but that in general, nebulization of a solution ispreferred over that of a suspension. He states that ultrasonicnebulizers should not be used on suspensions and should be used only onsolutions.

O'Callaghan (Thorax, (1990), 45, 109-111), Storr et al. (Arch. Dis.Child (1986), 61, 270-273), and Webb et al. (Arch. Dis. Child (1986),61, 1108-1110) suggest that nebulization of corticosteroid (inparticular beclomethasone) solutions may be preferred over that ofsuspensions because the latter may be inefficient if the nebulizedparticles are too large to enter the lung in therapeutically effectiveamounts. However, data presented by O'Callaghan (J. Pharm. Pharmacol.(2002), 54, 565-569) on the nebulization of flunisolide solution versussuspension showed that the two performed similarly. Therefore, it cannotbe generalized that nebulization of a solution is preferred over that ofa suspension.

Accordingly, there is a widely recognized need for a non-suspensionformulation comprising a corticosteroid for administration vianebulization. However, the PULMICORT® suspension unit dose formulationis widely available and accepted in the field of inhalation therapy. Itwould be of great benefit to this field of therapy to provide a methodof improving the administration of the PULMICORT® suspension unit doseformulation, or more generally, of a suspension unit dose formulationcontaining a corticosteroid.

However, the current focus in nebulizer therapy is to administer higherconcentrations of drug, use solution, preferably predominantlyaqueous-based solutions in preference to non-aqueous or alcoholic ornon-aqueous alcoholic solutions or suspensions if possible, minimizetreatment time, synchronize nebulization with inhalation, and administersmaller droplets for deeper lung deposition of drug.

Corticosteroid-containing solutions for nebulization are known. Thereare a number of different ways to prepare solutions for nebulization.These generally have been prepared by the addition of a cosolvent,surfactant, or buffer. However, cosolvents, such as ethanol,polyethylene glycol and propylene glycol are only tolerated in lowamounts when administered by inhalation due to irritation of therespiratory tract. There are limits to acceptable levels of thesecosolvents in inhaled products. Typically, the cosolvents make up lessthan about 35% by weight of the nebulized composition, although it isthe total dose of cosolvent as well as its concentration that determinesthese limits. The limits are set by the propensity of these solventseither to cause local irritation of lung tissue, to form hyperosmoticsolutions that would draw fluid into the lungs, and/or to intoxicate thepatient. In addition, most potential hydrophobic therapeutic agents arenot sufficiently soluble in these cosolvent mixtures.

Saidi et al. (U.S. Pat. No. 6,241,969) disclose the preparation ofcorticosteroid-containing solutions for nasal and pulmonary delivery.The dissolved corticosteroids are present in a concentrated, essentiallynon-aqueous form for storage or in a diluted, aqueous-based form foradministration.

Keller et al. (in Respiratory Drug Delivery IX (2004) 221-231) disclosethe deposition of solution formulations containing budesonide andsurfactants to children.

Lintz et al. (AAPS Annual Meeting and Exposition, Baltimore, Nov. 8,2004; Poster M1128) disclose the preparation and aerosolcharacterization of liquid formulations containing budesonide, water,citrate salt, sodium chloride and alcohol, propylene glycol and/orsurfactant, such as Tween, Pluronic, or phospholipids with HLB-valuesbetween 10 and 20. The aerosol characterization of such a combinationsurfactant solution in a vibrating membrane/mesh nebulizer (the ParieFlow) compared to a suspension was studied using adult and child breathsimulation. They reported a respirable fraction of 83.3% for the PARIeFlow with the solution and 61% for the PULMICORT RESPULES with the PARILC+. They also reported the results regarding the respirable drugdelivery rate (% drug <5 μm/min), delivered dose (% of drug charged todevice), drug delivery rate (% drug/min), and respirable dose (%<5 μm).

Schueepp et al. (ATS 99^(th) International Conference, Seattle, May16^(th)-21^(st), 2003; poster 1607) disclose assessment of the aerosolperformance of a customized eFlow Baby Functional Model with anexperimental budesonide solution (100 μg in 0.5 ml) utilizing a babycast model and applying different breathing patterns.

An alternative approach to administration of the PULMICORT™ suspensionis administration of a liposome formulation. Waldrep et al. (J. AerosolMed. (1994), 7(2), 135-145) reportedly succeeded in preparing a liposomeformulation of budesonide and phosphatidylcholine derivatives.

None of the above-identified formulations has provided a method ofimproving the administration of a suspension-based unit dose formulationcontaining a corticosteroid. Instead, the general focus of the art hasbeen to completely circumvent formulating a suspension by firstpreparing a liquid formulation that is then divided into multiple unitdoses that are packaged for marketing and then sold for use.

Solubilization of drugs by cyclodextrins and their derivatives is wellknown. Cyclodextrins are cyclic carbohydrates derived from starch. Theunmodified cyclodextrins differ by the number of glucopyranose unitsjoined together in the cylindrical structure. The parent cyclodextrinscontain 6, 7, or 8 glucopyranose units and are referred to as α-, β-,and γ-cyclodextrin respectively. Each cyclodextrin subunit has secondaryhydroxyl groups at the 2 and 3 positions and a primary hydroxyl group atthe 6-position. The cyclodextrins may be pictured as hollow truncatedcones with hydrophilic exterior surfaces and hydrophobic interiorcavities. In aqueous solutions, these hydrophobic cavities provide ahaven for hydrophobic organic compounds that can fit all or part oftheir structure into these cavities. This process, known as inclusioncomplexation, may result in increased apparent aqueous solubility andstability for the complexed drug. The complex is stabilized byhydrophobic interactions and does not involve the formation of anycovalent bonds.

This dynamic and reversible equilibrium process can be described byEquations 1 and 2, where the amount in the complexed form is a functionof the concentrations of the drug and cyclodextrin, and the equilibriumor binding constant, K_(b). When cyclodextrin formulations areadministered by injection into the blood stream, the complex rapidlydissociates due to the effects of dilution and non-specific binding ofthe drug to blood and tissue components.

$\begin{matrix}{{{Drug} + {Cyclodextrin}}\overset{K_{b}}{\leftrightarrow}{Complex}} & {{Equation}\mspace{14mu} 1} \\{K_{b} = \frac{\left\lfloor {Complex} \right\rfloor}{\left\lfloor {Drug} \right\rfloor \left\lfloor {Cyclodextrin} \right\rfloor}} & {{Equation}\mspace{14mu} 2}\end{matrix}$

Binding constants of cyclodextrin and an active agent can be determinedby the equilibrium solubility technique (T. Higuchi et al. in “Advancesin Analytical Chemistry and Instrumentation Vol. 4”; C. N. Reilly ed.;John Wiley & Sons, Inc, 1965, pp. 117-212). Generally, the higher theconcentration of cyclodextrin, the more the equilibrium process ofEquations 1 and 2 is shifted to the formation of more complex, meaningthat the concentration of free drug is generally decreased by increasingthe concentration of cyclodextrin in solution.

The underivatized parent cyclodextrins are known to interact with humantissues and extract cholesterol and other membrane components,particularly upon accumulation in the kidney tubule cells, leading totoxic and sometimes fatal renal effects.

The parent cyclodextrins often exhibit a differing affinity for anygiven substrate. For example, γ-cyclodextrin often forms complexes withlimited solubility, resulting in solubility curves of the type Bs. Thisbehavior is known for a large number of steroids which imposes seriouslimitations towards the use of γ-CD in liquid preparations. β-CD,however, does not complex well with a host of different classes ofcompounds. It has been shown for β-CD and γ-CD that derivatization, e.g.alkylation, results in not only better aqueous solubility of thederivatives compared to the parent CD, but also changes the type ofsolubility curves from the limiting Bs-type to the more linear A-typecurve (Bernd W. Muller and Ulrich Brauns, “Change of Phase-SolubilityBehavior by Gamma-Cyclodextrin Derivatization”, Pharmaceutical Research(1985) p 309-310).

Chemical modification of the parent cyclodextrins (usually at thehydroxyls) has resulted in derivatives with improved safety whileretaining or improving the complexation ability. Of the numerousderivatized cyclodextrins prepared to date, only two appear to becommercially viable: the 2-hydroxypropyl derivatives (HP-CD; neutral

Sulfobutyl Ether-β-Cyclodextrin (Captisol®)

cyclodextrins being commercially developed by Janssen and others), andthe sulfoalkyl ether derivatives, such as sulfobutyl ether, (SBE-CD;anionic cyclodextrins being developed by CyDex, Inc.) However, theHP-β-CD still possesses toxicity that the SBE-CD does not.

U.S. Pat. No. 5,376,645 and U.S. Pat. No. 5,134,127 to Stella et al.,U.S. Pat. No. 3,426,011 to Parmerter et al., Lammers et al. (Recl. Tray.Chim. Pays-Bas (1972), 91(6), 733-742); Staerke (1971), 23(5), 167-171)and Qu et al. (J. Inclusion Phenom. Macro. Chem., (2002), 43, 213-221)disclose sulfoalkyl ether derivatized cyclodextrins. The referencessuggest that SAE-CD should be suitable for solubilizing a range ofdifferent compounds. However, Stella discloses that the molar ratio ofsulfoalkyl ether derivatized cyclodextrin to active ingredient suitablefor solubilization of the active ingredient, even a corticosteroid, inwater ranges from 10:1 to 1:10.

A sulfobutyl ether derivative of beta cyclodextrin (SBE-β-CD), inparticular the derivative with an average of about 7 substituents percyclodextrin molecule (SBE7-β-CD), has been commercialized by CyDex,Inc. as CAPTISOL®. The anionic sulfobutyl ether substituent dramaticallyimproves the aqueous solubility of the parent cyclodextrin. In addition,the presence of the charges decreases the ability of the molecule tocomplex with cholesterol as compared to the hydroxypropyl derivative.Reversible, non-covalent, complexation of drugs with CAPTISOL®cyclodextrin generally allows for increased solubility and stability ofdrugs in aqueous solutions. While CAPTISOL® is a relatively new butknown cyclodextrin, its use in the preparation ofcorticosteroid-containing solutions for nebulization has not previouslybeen evaluated.

Hemolytic assays are generally used in the field of parenteralformulations to predict whether or not a particular formulation islikely to be unsuitable for injection into the bloodstream of a subject.If the formulation being tested induces a significant amount ofhemolysis, that formulation will generally be considered unsuitable foradministration to a subject. It is generally expected that a higherosmolality is associated with a higher hemolytic potential. As depictedin FIG. 1 (Thompson, D. O., Critical Reviews in Therapeutic Drug CarrierSystems, (1997), 14(1), 1-104), the hemolytic behavior of the CAPTISOL®is compared to the same for the parent β-cyclodextrin, the commerciallyavailable hydroxypropyl derivatives, ENCAPSIN™ cyclodextrin (degree ofsubstitution˜3-4) and MOLECUSOL® cyclodextrin (degree ofsubstitution˜7-8), and two other sulfobutyl ether derivatives, SBE1-β-CDand SBE4-β-CD. Unlike the other cyclodextrin derivatives, sulfoalkylether (SAE-CD) derivatives, in particular those such as the CAPTISOL(degree of substitution˜7) and SBE4-β-CD (degree of substitution˜4),show essentially no hemolytic behavior and exhibit substantially lowermembrane damaging potential than the commercially availablehydroxypropyl derivatives at concentrations typically used to solubilizepharmaceutical formulations. The range of concentrations depicted in thefigure includes the concentrations typically used to solubilizepharmaceutical formulations when initially diluted in the blood streamafter injection. After oral administration, SAE-CD does not undergosignificant systemic absorption.

The osmolality of a formulation is generally associated with itshemolytic potential: the higher the osmolality (or the more hypertonic),the greater the hemolytic potential. Zannou et al. (“Osmotic propertiesof sulfobutyl ether and hydroxypropyl cyclodextrins”, Pharma. Res.(2001), 18(8), 1226-1231) compared the osmolality of solutionscontaining SBE-CD and HP-CD. As depicted in FIG. 2, the SBE-CDcontaining solutions have a greater osmolality than HP-CD containingsolutions comprising similar concentrations of cyclodextrin derivative.Thus, it is surprising that SAE-CD exhibits lower hemolysis than doesHP-CD at equivalent concentrations, even though HP-CD has a lowerosmolality.

Methylated cyclodextrins have been prepared and their hemolytic effecton human erythrocytes has been evaluated. These cyclodextrins were foundto cause moderate to severe hemolysis (Jodal et al., Proc. 4^(th) Int.Symp. Cyclodextrins, (1988), 421-425; Yoshida et al., Int. J. Pharm.,(1988), 46(3), 217-222).

Administration of cyclodextrins into the lungs of a mammal may not beacceptable. In fact, literature exists on the potential or observedtoxicity of native cyclodextrins and cyclodextrin derivatives. The NTPChemical Repository indicates that α-cyclodextrin may be harmful byinhalation. Nimbalkar et al. (Biotechnol. Appl. Biochem. (2001), 33,123-125) cautions on the pulmonary use of an HP-β-CD/diacetyldapsonecomplex due to its initial effect of delaying cell growth of lung cells.

Even so, a number of studies regarding the use of cyclodextrins forinhalation have been reported although none have been commercialized.The studies suggest that different drug-cyclodextrin combinations willbe required for specific optimal or even useful inhaled or intra-nasalformulations. Attempts have been made to develop cyclodextrin-containingpowders and solutions for buccal, pulmonary and/or nasal delivery.

U.S. Pat. No. 5,914,122 to Otterbeck et al. discloses the preparation ofstable budesonide-containing solutions for rectal administration as afoam. They demonstrate the use of cyclodextrin, such as β-CD, γ-CD orHP-β-CD, and/or EDTA as a stabilizer. Cyclodextrin is also suggested asa solubilizer for increasing the concentration of budesonide insolution. In each case, the greatest shelf-life they report for any oftheir formulations is, in terms of acceptable retention of the activeingredient, only three to six months.

U.S. Pregrant Patent Publication No. 20020055496 to McCoy et al.discloses essentially non-aqueous intra-oral formulations containingHP-β-CD. The formulations may be administered with an aerosol, spraypump or propellant.

Russian Patent No. 2180217 to Chuchalin discloses a stablebudesonide-containing solution for inhalation. The solution comprisesbudesonide, propylene glycol, poly(ethylene oxide), succinic acid,Trilon B, nipazole, thiourea, water, and optionally HP-β-CD.

Müller et al. (Proceed. Int'l. Symp. Control. Rel. Bioact. Mater.(1997), 24, 69-70) discloses the results of a study on the preparationof budesonide microparticles by an ASES (Aerosol Solvent ExtractionSystem) supercritical carbon dioxide process for use in a dry powderinhaler. HP-β-CD is suggested as a carrier for a powder.

Müller et al. (U.S. Pat. No. 6,407,079) discloses pharmaceuticalcompositions containing HP-β-CD. They suggest that nasal administrationof a solution containing the cyclodextrin is possible.

The art recognizes that it may be necessary to evaluate structurallyrelated variations of a particular type of cyclodextrin derivative inorder to optimize the binding of a particular compound with that type ofcyclodextrin derivative. However, it is often the case that there arenot extreme differences in the binding of a particular compound with afirst embodiment versus a second embodiment of a particular cyclodextrinderivative. For example, cases where there are extreme differences inthe binding of a particular therapeutic agent for a first cyclodextrinderivative versus a structurally related second cyclodextrin derivativeare uncommon. When such situations do exist, they are unexpected. Worthet al. (24^(th) International Symposium on Controlled Release ofBioactive Materials (1997)) disclose the results of a study evaluatingthe utility of steroid/cyclodextrin complexes for pulmonary delivery. Inside-by-side comparisons, β-CD, SBE7-β-CD, and HP-β-CD were evaluatedaccording to their ability to form inclusion complexes withbeclomethasone dipropionate (BDP) and its active metabolitebeclomethasone monopropionate (BMP). BMP was more easily solubilizedwith a cyclodextrin, and the observed order of solubilizing power was:HP-β-CD (highest)>β-CD>SBE7-β-CD. Thus, the artisan would expect thatSAE-CD derivatives would not be as suitable for use in solubilizingcorticosteroids such as BMP or BDP. Although no results regarding actualutility in an inhaled formulation were disclosed, they suggest that BMPrather than BDP would be a better alternative for development of anebulizer solution.

Kinnarinen et al. (11^(th) International Cyclodextrin Symposium CD,(2002)) disclose the results of a study of the in vitro pulmonarydeposition of a budesonide/γ-CD inclusion complex for dry powderinhalation. No advantage was observed by complexation with γ-CD. Vozoneet al. (11^(th) International Cyclodextrin Symposium CD, (2002))disclose the results of a study on the complexation of budesonide withγ-cyclodextrin for use in dry powder inhalation. No difference wasobserved within emitted doses of the cyclodextrin complex or a physicalmixture of budesonide and the CD. But, a difference observed in the fineparticle fraction of both formulations suggested that use of acyclodextrin complex for pulmonary drug delivery might increase therespirable fraction of the dry powder.

Pinto et al. (S.T.P. Pharma. Sciences (1999), 9(3), 253-256) disclosethe results of a study on the use of HP-β-CD in an inhalable dry powderformulation for beclomethasone. The HP-β-CD was evaluated as a complexor physical mixture with the drug in a study of in vitro deposition ofthe emitted dose from a MICRO-HALER™ inhalation device. The amount ofrespirable drug fraction was reportedly highest with the complex andlowest with the micronized drug alone.

Rajewski et al. (J. Pharm. Sci. (1996), 85(11), 1142-1169) provide areview of the pharmaceutical applications of cyclodextrins. In thatreview, they cite studies evaluating the use of cyclodextrin complexesin dry powder inhalation systems.

Shao et al (Eur. J. Pharm. Biopharm. (1994), 40, 283-288) reported onthe effectiveness of cyclodextrins as pulmonary absorption promoters.The relative effectiveness of cyclodextrins in enhancing pulmonaryinsulin absorption, as measured by pharmacodynamics, and relativeefficiency was ranked as follows:dimethyl-β-cyclodextrin>α-cyclodextrin>β-cyclodextrin>γ-cyclodextrin>hydroxypropyl-β-cyclodextrin.In view of this report, the artisan would expect the water solublederivative of γ-CD to be less suitable for delivering compounds viainhalation than the respective derivative of β-CD because theunderivatized β-CD is more suitable than the underivatized γ-CD.

Williams et al. (Eur. J. Pharm. Biopharm. (1999 March), 47(2), 145-52)reported the results of a study to determine the influence of theformulation technique for 2-hydroxypropyl-beta-cyclodextrin (HP-β-CD) onthe stability of aspirin in a suspension-based pressurized metered-doseinhaler (pMDI) formulation containing a hydrofluoroalkane (HFA)propellant. HP-β-CD was formulated in a pMDI as a lyophilized inclusioncomplex or a physical mixture with aspirin. Aspirin in the lyophilizedinclusion complex exhibited the most significant degree of degradationduring the 6-months storage, while aspirin alone in the pMDIdemonstrated a moderate degree of degradation. Aspirin formulated in thephysical mixture displayed the least degree of degradation. Reportedly,HP-β-CD may be used to enhance the stability of a chemically labiledrug, but the drug stability may be affected by the method ofpreparation of the formulation.

Gudmundsdottir et al. (Pharmazie (2001 December), 56(12), 963-6)disclose the results of a study in which midazolam was formulated inaqueous sulfobutylether-beta-cyclodextrin buffer solution. The nasalspray was tested in healthy volunteers and compared to intravenousmidazolam in an open crossover trial. The nasal formulation reportedlyapproaches the intravenous form in speed of absorption, serumconcentration and clinical sedation effect. No serious side effects wereobserved.

Srichana et al. (Respir. Med. (2001 June), 95(6), 513-9) report theresults of a study to develop a new carrier in dry powder aerosols. Twotypes of cyclodextrin were chosen; gamma cyclodextrin (γ-CD) anddimethyl-beta-cyclodextrin (DMCD) as carriers in dry powderformulations. Salbutamol was used as a model drug and a controlformulation containing lactose and the drug was included. A twin-stageimpinger (TSI) was used to evaluate in delivery efficiency of those drypowder formulations. From the results obtained, it was found that theformulation containing γ-CD-enhanced drug delivery to the lower stage ofthe TSI (deposition=65%) much greater than that of both formulationscontaining DMCD (50%) and the control formulation (40%) (P<0.05). Thehaemolysis of red blood cells incubated with the DMCD complex was higherthan that obtained in the γ-CD complex. The drug release in bothformulations containing γ-CD and DMCD was fast (over 70% was released in5 min) and nearly all the drug was released within 30 min.

van der Kuy et al. (Eur. J. Clin. Pharmacol. (1999 November), 55(9),677-80) report the results of the pharmacokinetic properties of twointranasal preparations of dihydroergotamine mesylate (DHEM)-containingformulation using a commercially available intranasal preparation. Theformulations also contained randomly methylated β-cyclodextrin (RAMEB).No statistically significant differences were found in maximum plasmaconcentration (Cmax), time to reach Cmax (tmax), area under plasmaconcentration-time curve (AUC0-8 h), Frel(t=8 h) and Cmax/AUC(t=8 forthe three intranasal preparations. The results indicate that thepharmacokinetic properties of the intranasal preparations are notsignificantly different from the commercially available nasal spray.

U.S. Pat. Nos. 5,942,251 and 5,756,483 to Merkus cover pharmaceuticalcompositions for the intranasal administration of dihydroergotamine,apomorphine and morphine comprising one of these pharmacologicallyactive ingredients in combination with a cyclodextrin and/or adisaccharide and/or a polysaccharide and/or a sugar alcohol.

U.S. Pat. No. 5,955,454 discloses a pharmaceutical preparation suitablefor nasal administration containing a progestogen and a methylated(3-cyclodextrin having a degree of substitution of between 0.5 and 3.0.

U.S. Pat. No. 5,977,070 to Piazza et al. discloses a pharmaceuticalcomposition for the nasal delivery of compounds useful for treatingosteoporosis, comprising an effective amount of a physiologically activetruncated analog of PTH or PTHrp, or salt thereof and an absorptionenhancer selected from the group consisting of dimethyl-β-cyclodextrin.

U.S. Pat. No. 6,436,902 to Backstrom et al. discloses compositions andmethods for the pulmonary administration of a parathyroid hormone in theform of a dry powder suitable for inhalation in which at least 50% ofthe dry powder consists of (a) particles having a diameter of up to 10microns; or (b) agglomerates of such particles. A dry powder inhalerdevice contains a preparation consisting of a dry powder comprising (i)a parathyroid hormone (PTH), and (ii) a substance that enhances theabsorption of PTH in the lower respiratory tract, wherein at least 50%of (i) and (ii) consists of primary particles having a diameter of up to10 microns, and wherein the substance is selected from the groupconsisting of a salt of a fatty acid, a bile salt or derivative thereof,a phospholipid, and a cyclodextrin or derivative thereof.

U.S. Pat. No. 6,518,239 to Kuo et al. discloses a dispersible aerosolformulation comprising an active agent and a dipeptide or tripeptide foraerosolized administration to the lung. The compositions reportedly mayalso include polymeric excipients/additives, e.g.,polyvinylpyrrolidones, derivatized celluloses such ashydroxymethylcellulose, hydroxyethylcellulose, and hydroxypropylmethylcellulose, Ficolls (a polymeric sugar), hydroxyethylstarch,dextrates (e.g., cyclodextrins, such as 2-hydroxypropyl-β-cyclodextrinand sulfobutylether-β-cyclodextrin), polyethylene glycols, and pectin.

Nakate et al. (Eur. J. Pharm. Biopharm. (2003 March), 55(2), 147-54)disclose the results of a study to determine the improvement ofpulmonary absorption of the cyclopeptide FK224 (low aqueous solubility)in rats by co-formulating it with beta-cyclodextrin. The purpose of thestudy was to investigate the effect of pulmonary delivery on thesystemic absorption of FK224 in comparison with other administrationroutes, and to determine the bioavailability (BA) of FK224 followingpulmonary administration in rats using various dosage forms. Afteradministration of an aqueous suspension, the bioavailability was reducedto 2.7% compared with 16.8% for the solution. However, β-cyclodextrin(β-CD) was found to be an effective additive as far as improving thesolubility of FK224 was concerned. The bioavailability of the aqueoussuspension containing β-CD was increased to 19.2%. It was observed thatboth the C(max) and AUC of FK224 were increased as the amount of β-CDincreased. The plasma profiles showed sustained absorption. They suggestthat β-CD is an extremely effective additive as far as improving thepulmonary absorption of FK224 is concerned. They also suggest that β-CDor derivatives with various degrees of aqueous solubility are potentialdrug carriers for controlling pulmonary absorption.

Kobayashi et al. (Pharm. Res. (1996 January), 13(1), 80-3) disclose theresults of a study on pulmonary delivery of salmon calcitonin (sCT) drypowders containing absorption enhancers in rats. After intratrachealadministration of sCT dry powder and liquid (solution) preparations torats, plasma sCT levels and calcium levels were measured. Reportedly,sCT in the dry powder and in the liquid were absorbed nearly to the samedegree. Absorption enhancers (oleic acid, lecithin, citric acid,taurocholic acid, dimethyl-β-cyclodextrin, octyl-β-D-glucoside) weremuch more effective in the dry powder than in the solution.

Adjei et al. (Pharm. Res. (1992 February), 9(2), 244-9) disclose theresults of a study on the bioavailability of leuprolide acetatefollowing nasal and inhalation delivery to rats and healthy humans.Systemic delivery of leuprolide acetate, a luteinizing hormone releasinghormone (LHRH) agonist, was compared after inhalation (i.h.) andintranasal (i.n.) administration. The i.n. bioavailability in rats wassignificantly increased by α-cyclodextrin (CD), EDTA, and solutionvolume. Absorption ranged from 8 to 46% compared to i.v. controls.Studies in healthy human males were conducted with leuprolide acetatei.n. by spray, or inhalation aerosol (i.h.), and subcutaneous (s.c.) andintravenous (i.v.) injection. The s.c. injection was 94% bioavailablecompared with i.v. The i.n. bioavailability averaged 2.4%, withsignificant subject-to-subject variability. Inhalation delivery gave aslightly lower intersubject variability. Mean Cmaxwith a 1-mg dose ofsolution aerosol was 0.97 ng/ml, compared with 4.4 and 11.4 ng/ml forsuspension aerosols given at 1- and 2-mg bolus dosages, respectively.The mean bioavailability of the suspension aerosols (28% relative tos.c. administration) was fourfold greater than that of the solutionaerosol (6.6%).

CyDex (Cyclopedia (2002), 5(1), 3) discloses that SBE-CD is non-toxic torats in an inhaled aerosol composition when present alone. They do notdisclose a nebulizable composition comprising a drug, in particular acorticosteroid, and SBE-CD. Studies comparing the nebulization ofPULMICORT RESPULES and an aqueous inhalable solution of budesonide andSAE-CD in various nebulizers have been disclosed (Zimmerer et al.Respiratory Drug Delivery IX (2004) 461-464).

In deciding whether to administer a suspension versus solution, one mustalso consider the type of nebulizer to be used. The two most commontypes of nebulizers are the ultrasonic nebulizer and the air driven jetnebulizer. There are significant differences between the two. Forexample, jet nebulizers cool rather than heat the liquid in thereservoir, whereas ultrasonic nebulizers heat the liquid. While heatingof the solution in reservoir can reduce the viscosity of the solutionand enhance formation of droplets, excessive heating could lead to drugdegradation. The ultrasonic nebulizer is quieter and provides fasterdelivery than the jet nebulizer, but ultrasonic nebulizers are moreexpensive and are not advised for the administration of the currentlyavailable steroid for nebulization. Most importantly, however,ultrasonic nebulizers generally provide a significantly higher rate ofadministration than do jet nebulizers.

Patients with asthma are often treated with inhaled short acting or longacting β2-agonists, inhaled anticholinergics, and inhaledcorticosteroids alone, sequentially or in combination. Combinations ofinhaled corticosteroids and long acting β2-agonists are known, forexample budesonide plus formoterol or fluticasone plus salmeterol areavailable in a dry powder inhaler. However, there is no example of suchcombinations that are available as a solution for nebulization.Combining the medications into one solution would reduce the timerequired to administer the medications separately.

For inhaled corticosteroids, high pulmonary availability is moreimportant than high oral bioavailability because the lung is the targetorgan. A product with high pulmonary availability has greater potentialto exert positive effects in the lung. The ideal ICS would have minimumoral bioavailability, reducing the likelihood of systemic adverseeffects.

Although extremely effective in the treatment of asthma, inhaledcorticosteroids can have a number of adverse side effects such as oralcandidiasis, hoarseness (dysphonia), and pharyngitis. Therefore, inhaledcorticosteroids are best delivered by a method that minimizes the oraland/or pharyngeal deposition of the corticosteroid and instead maximizespulmonary delivery.

Some corticosteroids posses a hydroxyl group at position 21 of thecorticosteroid. Those compounds include budesonide, flunisolide,triamcinolone acetonide, beclomethasone monopropionate, and the activeform of ciclesonide (desisobutyryl-ciclesonide). It is known thatciclesonide is inhaled as an inactive compound and converted byesterases in the lung to its active form, desisobutyryl-ciclesonide(des-CIC). Budesonide conjugates to form intracellular fatty acidesters, which are highly lipophilic. Budesonide forms conjugates with 5fatty acids: oleate, palmitate, linoleate, palmitoleate, andarachidonate.

In summary, the art suggests that, in some cases, nebulization ofsolutions may be preferred over that of suspensions and that, in somecases, an ultrasonic nebulizer, vibrating mesh, electronic or othermechanism of aerosolization may be preferred over an air driven jetnebulizer depending upon the nebulization liquid formulations beingcompared. Even though the art discloses inhalable solution formulationscontaining a corticosteroid and cyclodextrin, the results of the art areunpredictable. In other words, the combination of one cyclodextrin withone drug does not suggest that another cyclodextrin may be suitable.Neither does the art suggest that one cyclodextrin-corticosteroidinhalable formulation will possess advantages over anothercyclodextrin-corticosteroid inhalable formulation.

A need remains in the art for a stabilized aqueous solutionbudesonide-containing inhalable formulation that does not require theaddition of preservatives and that provides significant advantages overother stabilized aqueous solution budesonide-containing inhalableformulations. A need also remains for a method of improving theadministration of budesonide-containing suspension formulations bynebulization by converting the suspension to a solution.

There is also a need to develop improved systems that can solubilizewater-insoluble drugs for nebulization, and to minimize the levels ofcosolvent necessary to accomplish this. The ideal system would consistof non-toxic ingredients and be stable for long periods of storage atroom temperature. When nebulized, it would produce respirable dropletsin the less than 10 micron or less than 5 micron or less than 3 micronand a substantial portion of extra-fine aerosol in the less than about 1micron size range.

The need continues to remain for a method of improving theadministration, by nebulization, of a suspension-based unit doseformulation. Such a method would reduce the overall time ofadministration, increase the overall amount of drug administered, reducethe amount of drug left in the reservoir of the nebulizer, increase theportion of pulmonary deposition relative to oropharyngeal deposition ofcorticosteroid, and/or enhance deep lung penetration of thecorticosteroid as compared to such administration, absent theimprovement, of the suspension-based unit dose formulation.

SUMMARY OF THE INVENTION

The present invention seeks to overcome the disadvantages present inknown formulations. As such, a derivatized cyclodextrin-based, e.g.,sulfoalkyl ether cyclodextrin (SAE-CD)-based, inhalable formulation isprovided. The present formulation includes a corticosteroid as aprinciple active agent. The present formulation may provide enhancedsolubility and/or enhanced chemical, thermochemical, hydrolytic and/orphotochemical stability of the active agent or other ingredients in theformulation. Moreover, the present formulation may possess otheradvantages, e.g. enhanced drug delivery, increased rate of drugadministration, reduced treatment time, reduced toxicity, ease ofmanufacture, assurance of sterility, improved stability, enhancedbioabsorption, no requirement of particle size control, increased outputrate, increased total output, no concern for solid particle growth,enhanced pharmacokinetic profile, reduced corticosteroid-related sideeffects, enhanced pulmonary deposition, reduced oropharyngealdeposition, improved nebulization performance, and/or no need to confirmformation of a suspension, over other inhalable solution or suspensionformulations containing a corticosteroid, such as budesonide.

The invention provides a formulation, system and method for treating arespiratory disease or disorder that is therapeutically responsive tocorticosteroid therapy. The system of the invention comprises anebulizer, and a liquid formulation of the invention. The liquidformulation of the invention comprises a dose of corticosteroid, anaqueous liquid carrier and a solubility enhancer.

The invention provides a method of treating, preventing or amelioratingin a subject a corticosteroid-responsive disease or disorder, meaning adisease or disorder in a subject that can be treated with atherapeutically effective amount of corticosteroid to provide a clinicalor therapeutic benefit to the subject. The method comprises: providingan aqueous inhalable formulation comprising a corticosteroid, an aqueousliquid carrier, and solubility enhancer; and delivering or administeringvia nebulization to a subject in need thereof a dose of about at least25 μg, 40 μg, at least 45 μg, at least 48 μg, 45-1000 μg, about 1 μg to20 mg, or 1 μg to 10 mg, 0.01 mg to 10 mg, 0.025 mg to 10 mg, 0.05 mg to5 mg, 0.1 mg to 5 mg, 0.125 mg to 5 mg, 0.25 mg to 5 mg, 0.5 mg to 5 mg,0.05 mg to 2 mg, 0.1 mg to 2 mg, 0.125 mg to 2 mg, 0.25 mg to 2 mg, 0.5mg to 2 mg, 1 μg, 10 μg, 25 μg, 50 μg, 100 μg, 125 μg, 200 μg, 250 μg,25 to 66 μg, 48 to 81 μg, 73 to 125 μg, 40 μg, 64 μg, 95 μg, 35 to 95μg, 25 to 125 μg, 60 to 170 μg, 110 μg, 170 μg, 45 to 220 μg, 45 to 85μg, 48 to 82 μg, 85 to 160 μg, 140 to 220 μg, 120 to 325 μg, 205 μg, 320μg, 325 μg, 90 to 400 μg, 95 to 170 μg, 165 to 275 μg, or 275 to 400 μgof the corticosteroid. In some embodiments, the corticosteroid isbudesonide.

In some embodiments: 1) the inhalable formulation has a volume of about10 p. 1 to 100 ml, 50 μl to 50 ml, 50 μl to 10 ml, 0.1 to 10 ml, 0.1 mlto less than 10 ml, 0.1 ml to 7.5 ml, 0.1 ml to 5 ml, 0.1 ml to 3 ml,0.1 ml to 2 ml, 0.1 ml to 1 ml, 0.05 ml to 7.5 ml, 0.05 ml to 5 ml, 0.05ml to 3 ml, 0.05 ml to 2 ml, or 0.05 ml to 1 ml; 2) the step ofdelivering or administering is conducted over a period of less than 30min, less than 20 min, less than 10 min, less than 7 min, less than 5min, less than 3 min, or less than 2 min, or the time is about 0.05 to10 min, about 0.1 to 5 min, about 0.1 to 3 min, about 0.1 to 2 min,about 0.1 to 1.5 min, about 0.5 min to about 1.5 min, or about 1 min, orthe time is about the time it takes for a subject to take a singlebreath (about 1 to 3 or 1 to 5 sec); 3) 20% to 85%, or 30% to 80%, or atleast 30%, at least 40%, at least 56%, at least 59%, at least 62% of thedose of corticosteroid is delivered to the lungs of the subject; 4) theformulation, system or method provides an enhanced pharmacokineticprofile of 1.5 to 8 fold, 1.5 to 6 fold, 1.5 to 4 fold, 1.5 to 2 fold,1.64 to 3.55 fold for Cmax, 1.48 to 6.25 fold for Cmax, 1.59 to 3.55 or1.19 to 6.11 fold for AUC_(inf), 1.69-3.67 or 1.21 to 7.66 fold forAUC_(last), on normalized dose, as compared to administration of asuspension formulation comprising approximately the same amount ofcorticosteroid and delivered under substantially the same conditions; 5)no more than, 15% to 85%, 15% to 70%, no more than 20% to 70%, no morethan 50%, no more than 40%, or no more than 60% of the dose ofcorticosteroid is delivered to the subject outside the lung.

The present inventors have unexpectedly discovered that SAE-CD issystemically absorbed following administration via inhalation. It isalso eliminated from the lungs. SAE-CD also complexes withcorticosteroids in aqueous inhalable liquid formulations.Coadministration of the corticosteroid with SAE-CD may result inincreased output rate and total drug delivery as compared to a controlexcluding SAE-CD. Moreover, an inhalable solution of the inventionadministered by nebulization provides other advantages set forth hereinas compared to a control/reference sample of a suspension comprisingsubstantially the same amount of corticosteroid in substantially thesame volume of liquid when administered under substantially the sameconditions.

The inhalable solution can be used in more types of nebulizers assuspensions can clog vibrating mesh nebulizers and do not perform wellin ultrasonic nebulizers. The nebulizers would be more easily cleaned ofresidual corticosteroid resulting in less contamination. As drug insolution will be distributed equally in the large and small dropletsleaving the nebulizer, the fine particle fraction will contain morecorticosteroid resulting in a greater respirable dose that can reach theperipheral lung and smaller airways. This may result in less of thetotal drug depositing in the oropharyngeal cavity and being swallowed,thus reducing the potential for undesired side effects while enhancingbioavailability and maximizing the desired effect. The inhalablesolution will also allow reducing the amount of drug in the unit doseand/or reducing the time of nebulization to achieve the desired amountof drug in the lung. The inhalable solution is more easily manufacturedand filled into unit doses and product performance does not depend onvery tight control of the particle size of ingoing corticosteroid andmaintenance of a homogeneous suspension during filling. However, smallerparticle size will, in general, facilitate more rapid dissolution duringmanufacturing. It is not necessary to maintain tight control asdescribed above. It may be terminally sterilized without risk ofchanging the corticosteroid's particle size of the suspendedcorticosteroid. The inhalable solution is not subject to change due toparticle growth via Ostwald ripening or agglomeration thus the amount ofdrug in the respirable fraction will not change over time. It does notneed to be resuspended prior to administration with carefully scriptedprocedures to reduce foaming so as not to create large variation isdose. It may be more accurately diluted to achieve the desired amount ofdrag in the volume placed in the nebulizer. Alternatively, doseadjustment may be controlled more easily by adjusting the time ofnebulization as the concentration of the inhalable solution does notchange as fast in the nebulizer as does concentration of the suspension,especially in air-jet nebulizers.

An SAE-CD-containing formulation can be prepared with sufficient activeagent solubility and stability for a commercial product. If needed, theSAE-CD-containing formulation can be prepared as a clear aqueoussolution that can be sterile filtered through a filter having a poresize of 0.45 μm or less and that is stable and preserved under a varietyof storage conditions. The invention thus provides afiltration-sterilized liquid formulation comprising a solution of theinvention and a method of sterilizing a solution of the invention bysterile filtration through a filter. Sterile filtration can be donewithout substantial mass loss of solubilized corticosteroid, meaningless than 5% mass loss.

Any corticosteroid suitable for administration via inhalation can beused according to the invention. Exemplary suitable corticosteroids arelisted herein. Some embodiments of the invention comprise acorticosteroid having a lipophilicity approximating or exceeding that offlunisolide. Some embodiments of the invention exclude a corticosteroidhaving a lipophilicity less than flunisolide, i.e, embodiments excludinghydrocortisone, prednisolone, prednisone, dexamethasone, betamethasone,methylprednisolone, triamcinolone, fluocortolone.

One aspect of the invention provides a liquid formulation comprising aneffective amount of corticosteroid, such as budesonide, and an SAE-CD,wherein the SAE-CD is present in an amount sufficient to dissolve andstabilize the corticosteroid during storage.

Another aspect of the invention provides a method of improving theadministration of corticosteroid to a subject by nebulization, themethod comprising the steps of:

providing in a unit dose an aqueous suspension formulation comprisingwater and corticosteroid suspended therein;

combining the suspension with an amount of SAE-CD sufficient to and fora period of time sufficient to solubilize the corticosteroid and form asolution; and

administering the solution to the subject, wherein the amount of timerequired to administer a therapeutic dose of corticosteroid with thesolution is less than the amount of time required to administer the sametherapeutic dose of corticosteroid with the suspension under similar, orotherwise comparable, nebulization conditions.

When administered with a nebulizer, a suspension for nebulization willprovide a first corticosteroid output rate under a first set ofnebulization conditions. However, when SAE-CD is added to the suspensionand mixed therein, a sufficient amount of the corticosteroid isdissolved to form a liquid formulation for nebulization that provides agreater corticosteroid output rate as compared to the formulationexcluding the SAE-CD when administered under substantially the sameconditions. In one embodiment, the drug output rate of the formulationis increased over that of the suspension even though the total volume ofnebulized composition, i.e., the total volume of solution emitted by thenebulizer, has not increased. In another embodiment, SAE-CD is presentin an amount sufficient to solubilize at least 50%, at least 75%, atleast 90%, at least 95% or substantially all of the corticosteroid. Someembodiments of the invention include those wherein at least 50% wt., atleast 75% wt., at least 90% wt., at least 95% wt., at least 98% wt., orall of the corticosteroid is dissolved in the inhalable liquidformulation.

In some embodiments, SAE-CD is present in an amount sufficient todecrease the amount of unsolubilized corticosteroid in the suspensionformulation and to improve the administration of the suspensionformulation via nebulization. In some embodiments, SAE-CD is present inan amount sufficient to solubilize enough corticosteroid such that thesuspension formulation to which the SAE-CD was added is converted to asolution, substantially clear solution (containing less than 5%precipitate or solid), or a clear solution. It is possible that othercomponents of the suspension formulation will not completely dissolvein, or may separate out from, the solution formulation containingSAE-CD.

According to some embodiments, a nebulizer charged with acorticosteroid/SAE-CD-containing solution generates smaller dropletsthan does the same nebulizer charged with acorticosteroid/HP-β-CD-containing solution operated under otherwisesimilar conditions. As a result of generating smaller droplets, thesystem comprising SAE-CD is improved over an otherwise similar systemcomprising HP-β-CD, since the SAE-CD based system will generate agreater proportion of respirable droplets and permit deeper lungpenetration.

Some aspects of the invention provides for the use of SAE-CD in anebulizable unit dose liquid formulation. In one embodiment, theinvention provides use of SAE-CD for converting a nebulizablecorticosteroid-containing suspension-based unit dose formulation to anebulizable corticosteroid-containing liquid unit dose formulation.

Some embodiments of the invention include those wherein: 1) thecorticosteroid to SAE-CD molar ratio is 0.5 to 0.0001 (1:2 to 1:10,000),1:1 to 1:100, 1:1 to 1:10,000, or 0.1 (1:10) to 0.03 (1:33.33). Themolar ratio of SAE-CD to corticosteroid in the formulation or system isgenerally greater than 10:1, greater than about 11:1, greater than 13:1,or greater than 14:1. Depending upon the corticosteroid used in theformulation, the molar ratio of corticosteroid to SAE-CD can vary inorder to obtain a solution suitable for administration via inhalationfor the treatment of a respiratory disease or disorder. In someembodiments, the nebulization composition comprises at least 4.8±0.5%wt./vol of SAE-CD to provide a self-preserved formulation for a periodpredetermined period of time. In some embodiments, the nebulizationcomposition comprises less than or about 21.5±2% wt./wt. of SAE-CD. Insome embodiments, the SAE-CD is present in an amount sufficient toprovide a clear solution. For example, the nebulization composition canbe visibly clear as viewed by the unaided eye.

Some suitable SAE-CD's include, for example, sulfobutyl ether 4-β-CD orsulfobutyl ether 7-β-CD, sulfobutyl ether 6-γ-CD, sulfobutyl ether4-γ-CD, sulfobutyl ether 3 to 8-γ-CD, or a sulfobutyl ether 5-γ-CD, or acompound of the formula 1 or a mixture thereof.

A composition of the invention can further comprise a conventionalpreservative, an antioxidant, a buffering agent, an acidifying agent, asolubilizing agent, a complexation enhancing agent, saline, anelectrolyte, another therapeutic agent, an alkalizing agent, a tonicitymodifier, surface tension modifier, viscosity modifier, surfactant,density modifier, volatility modifier, or a combination thereof. Ifdesired, the composition further comprises a liquid carrier other thanwater. If a conventional preservative is included in the composition,the corticosteroid, such as budesonide, may have a greater binding withthe SAE-CD than does a conventional preservative. A composition can bepurged with an inert gas prior to storage to remove substantially all ofthe oxygen contained in the formulation. In general, the formulation orcomposition of the invention has a shelf-life of at least 6 monthsdepending upon the intended use.

The formulation can be prepared at a temperature at or above 5° C., ator above 25° C., at or above 35° C., at or above 45° C. or at or above50° C. Specific embodiments of the methods of preparing a liquidformulation include those wherein: 1) the method further comprises thestep of sterile filtering the formulation through a filtration mediumhaving a pore size of 0.1 microns or larger; 2) the liquid formulationis sterilized by irradiation or autoclaving; 3) the nebulizationsolution is purged with nitrogen or argon or other inertpharmaceutically acceptable gas prior to storage such that a substantialportion of the oxygen dissolved in, and/or in surface contact with thesolution is removed.

The invention provides a method of stabilizing corticosteroid in anaqueous corticosteroid-containing formulation comprising the step ofadding SAE-CD to an aqueous corticosteroid-containing suspension ofsolution formulation in an amount sufficient to reduce the rate ofdegradation of corticosteroid as compared to a control sample excludingSAE-CD.

The invention also provides a method of improving the administration ofan inhalable aqueous corticosteroid-containing suspension unit doseformulation by nebulization, the method comprising the step of addingSAE-CD to an aqueous corticosteroid-containing suspension unit doseformulation in an amount sufficient to solubilize the corticosteroid toform an inhalable aqueous corticosteroid-containing solution unit doseformulation, the improvement comprising increasing the output rateand/or extent of nebulized corticosteroid.

The invention provides a method of reducing the amount of time requiredto provide a therapeutically effective amount of corticosteroid to asubject by inhalation of an corticosteroid-containing composition with anebulizer, the method comprising the steps of: including SAE-CD in thecomposition in an amount sufficient to solubilize the corticosteroid toform an inhalable aqueous corticosteroid-containing solution; andadministering the solution to the subject by inhalation with anebulizer, wherein the amount of time required to provide atherapeutically effective amount of corticosteroid to the subject withthe solution is reduced as compared to the amount of time required toprovide a therapeutically effective amount of corticosteroid to thesubject with a corticosteroid-containing suspension comprising the sameamount or concentration of corticosteroid when the suspension andsolution are administered under otherwise similar nebulizationconditions. In some embodiments, the time required to administer ordeliver a dose of corticosteroid is less than 30 min, less than 20 min,less than 10 min, less than 7 min, less than 5 min, less than 3 min, orless than 2 min, or the time is about 0.05 to 10 min, about 0.1 to 5min, about 0.1 to 3 min, about 0.1 to 2 min, about 0.1 to 1.5 min, about0.5 min to about 1.5 min, or about 1 min, or about the time it takes fora subject to take a single breath (about 1 to 3 sec, or 1 to 5 sec). Thetime will vary according to the dose of corticosteroid in, theconcentration of corticosteroid in, and the volume of the inhalablecomposition in the reservoir of a nebulizer, and it will also dependupon the nebulizer format, nebulization efficiency, and reservoirvolume. In a given nebulizer, the lower the volume of the inhalablecomposition, the more quickly a corresponding dose of corticosteroid isnebulized. The higher the concentration of corticosteroid in aninhalable composition, the faster a dose of the corticosteroid can beadministered or delivered.

The output rate (the rate at which the dose corticosteroid isadministered or delivered) will vary according to the performanceparameters of the nebulizer used to administer it. The higher the outputrate of a given nebulizer, the lower the amount of time required todeliver or administer an inhalable composition, as defined herein, ordose of corticosteroid.

The invention also provides an inhalable composition comprising a watersoluble γ-CD derivative, a corticosteroid (either esterified orunesterified) and an aqueous liquid medium. Another embodiment of theinvention also provides an inhalable composition comprising a watersoluble β-CD derivative, a corticosteroid (unesterified) and an aqueousliquid medium.

Also, the invention provides an improved system for administering acorticosteroid-containing inhalable formulation by inhalation, theimprovement comprising including SAE-CD in the inhalable formulationsuch that SAE-CD is present in an amount sufficient to provide anincreased rate of inhaled corticosteroid as compared to administrationof a control inhalable formulation excluding SAE-CD but otherwise beingadministered under approximately the same conditions.

The invention can be used to provide a system for administration of acorticosteroid by inhalation, the system comprising an inhalationdevice, such as a nebulizer, and a drug composition comprising atherapeutically effective amount of corticosteroid, liquid carrier andSAE-CD present in an amount sufficient to solubilize the corticosteroidwhen presented to an aqueous environment, wherein the molar ratio ofcorticosteroid to SAE-CD is in the range of about 0.072 (1:13.89 orabout 1:14) to 0.0001 (1:10,000) or 0.063 (1:15.873 or about 1:16) to0.003 (1:333.33 or about 1:333). The molar ratio of SAE-CD tocorticosteroid ranges from >10:1 to about 1000:1, about from >10:1 toabout 100:1, from >10:1 to about 50:1, from >10:1 to about 30:1,from >10:1 to about 500:1. During operation, the system forms dropletshaving a MMAD in the range of about 1-8μ or 3-8μ. The corticosteroid isdelivered at a rate, of at least about 20-50 μg/min, wherein this rangemay increase or decrease according to the concentration ofcorticosteroid in the nebulization solution in the reservoir of thenebulizer.

As a result of using SAE-CD corticosteroid therapy with an inhalablenebulization solution, one can expect advantages such as enhanced drugdelivery, enhanced delivery especially to the peripheral or smallairways facilitated by the finer aerosol produced, potentially improvedtreatment of nocturnal, asymptomatic asthma and recovery from acuteasthma attacks, increased rate of drug administration, enhancedpharmacokinetic profile, reduced treatment time, improved formulationstability and/or improved patient compliance as compared to comparablecorticosteroid therapy with an inhalable nebulization suspension orsuspension chlorofluorocarbon (CFC) or hydrofluoroalkanes (HFA)pressurized metered-dose inhaler (pMDI).

The invention can be employed in a kit comprising SAE-CD, an aqueouscarrier, and corticosteroid, wherein the kit is adapted for thepreparation of a nebulization solution. Embodiments of the kit aredetailed below. The invention provides the potential to accommodatecombination products to overcome incompatibilities with suspension byother solution dosage forms.

The liquid formulation and composition of the invention provide anenhanced bioavailability/bioabsorption of the corticosteroid as comparedto a suspension-based aqueous formulation containing approximately thesame amount of corticosteroid and excluding SAE-CD. The liquidformulation or composition provides a higher ratio of AUCi (or AUCt) toamount of budesonide delivered to a subject than does the suspensionbased formulation. Thus, the liquid formulation and/or compositionprovides an improved clinical benefit or therapeutic benefit over thesuspension-based formulation.

The invention also provides a method of administering a corticosteroidto a subject comprising the step of administering via inhalation to asubject an aqueous liquid composition comprising a unit dose ofcorticosteroid dissolved therein, wherein the composition provides amean AUCt, normalized for dose of the corticosteroid, of at least 6(pg*h/ml) per μg of corticosteroid administered. In some embodiments,the unit dose comprises at least 45 μg, at least 48 μg, 45-1000 μg,about 1 μg to 20 mg, about 1 μg to 10 mg, 0.01 mg to 10 mg, 0.025 mg to10 mg, 0.05 mg to 5 mg, 0.1 mg to 5 mg, 0.125 mg to 5 mg, 0.25 mg to 5mg, 0.5 mg to 5 mg, 0.05 mg to 2 mg, 0.1 mg to 2 mg, 0.125 mg to 2 mg,0.25 mg to 2 mg, 0.5 mg to 2 mg, 1 μg, 10 μg, 25 μg, 50 μg, 100 μg, 125μg, 200 μg, 250 μg, 25 to 66 μg, 48 to 81 μg, 73 to 125 μg, 40 μg, 64μg, 95 μg, 35 to 95 μg, 25 to 125 μg, 60 to 170 μg, 110 μg, 170 μg, 45to 220 μg, 45 to 85 μg, 48 to 82 μg, 85 to 160 μg, 140 to 220 μg, 120 to325 μg, 205 μg, 320 μg, 325 μg, 90 to 400 μg, 95 to 170 μg, 165 to 275μg, or 275 to 400 μg of corticosteroid. In some embodiments, thenormalized AUCt is based upon the dose to subject, the normalized AUCtis based upon the dose to lung of subject, or the normalized AUCt isbased upon the emitted dose or nominal dose. As used herein, AUCt (orAUC_(last), AUC_(0-t)) is the area under the plasma level versus timecurve to the last time point t for which there is a quantifiable value.

The invention also provides a method of administering a corticosteroidto a subject comprising the step of administering via inhalation to asubject an aqueous liquid composition comprising a unit dose ofcorticosteroid dissolved therein, wherein the composition provides amean AUCi, normalized for dose of the corticosteroid, of at least 8(pg*h/ml) per μg of corticosteroid administered. In some embodiments,the unit dose comprises at least 45 μg, at least 48 μg, or 45-1000 μg,about 1 μg to 20 mg, about 1 μg to 10 mg, 0.01 mg to 10 mg, 0.025 mg to10 mg, 0.05 mg to 5 mg, 0.1 mg to 5 mg, 0.125 mg to 5 mg, 0.25 mg to 5mg, 0.5 mg to 5 mg, 0.05 mg to 2 mg, 0.1 mg to 2 mg, 0.125 mg to 2 mg,0.25 mg to 2 mg, or 0.5 mg to 2 mg of corticosteroid. In someembodiments, the normalized AUCi is based upon the dose to subject, thenormalized AUCi is based upon the dose to lung of subject, or thenormalized AUCi is based upon the emitted dose or nominal dose. As usedherein, the AUCi (AUC_(inf), AUC_((0→∞)) is the AUC extrapolated toinfinity.

When the aqueous liquid solution formulation of the invention isadministered via inhalation it can provide at least a two-fold increasein the AUCt or AUCi, normalized for dose of the corticosteroid, permicrogram of corticosteroid administered as compared to administrationvia inhalation of an aqueous liquid suspension formulation comprisingsubstantially the same amount of corticosteroid suspended therein. Insome embodiments, the formulation provides at least a 1.6 to five-fold(range as determined on an individual subject basis or a mean of 3 witha standard deviation of +/−1 as determined from the individual subjectdata) or a two to four-fold increase (as determined using the geometricmean across a patient population) in the AUCt or AUCi, normalized fordose of the corticosteroid, per microgram of corticosteroidadministered. The basis for amount of “corticosteroid administered” canbe “dose to subject” or “dose to lung” or “emitted dose” or “nominaldose” or “nominal available dose” or “loaded dose”.

The aqueous liquid formulation can be administered with any type ofdevice suitable for administration of a liquid formulation to the lung.For example, the formulation can be administered with an air driven jet,ultrasonic, capillary, electromagnetic, pulsating membrane, pulsatingplate (disc), pulsating mesh nebulizer, a nebulizer comprising avibration generator and an aqueous chamber, and a nebulizer comprising anozzle array.

In some embodiments, the invention provides a system comprising anebulizer and an aqueous liquid formulation/composition as describedherein. When charged with a liquid formulation of the inventioncomprising a dose of corticosteroid, the nebulizer can provide adecreasing rate of increasing concentration (RIC) of corticosteroid inthe formulation in the reservoir of the nebulizer, in particular whencomparing the performance of the liquid formulation to the performanceof a suspension-based formulation, e.g. PULMICORT RESPULES, in thenebulizer. In some embodiments, the percentage decrease in the RICranges from 10% to 60%, 15% to 60%, 20% to 60%, 30% to 60%, or 40% to60%. In some embodiments, the system may provide a RIC of 0 to 40, 1 to40, 5 to 30, or 10 to 30 mcg of corticosteroid/mL of solution volume permin of operation/nebulization.

The pharmacokinetic enhancement provided by the system andformulation/composition of the invention can be determined by comparisonof Cmax, AUC and Tmax parameters. In some embodiments, the formulationand system of the invention can provide a C_(max) (pg ofcorticosteroid/ml) of 90 to 900, 200 to 600, 200 to 550, 200 to 250, 400to 450, 500 to 600, 225, 437, or 545 on a dose non-normalized basis. Theformulation and system of the invention can provide a dose normalizedC_(max) (pg/ml/μg) of: 1) 0.3 to 2, 0.35 to 2, 0.6 to 1.5, 0.5 to 1.2,0.8 to 1, 0.8 to 0.9, 0.7 to 0.8, 0.4, 1.9, 0.6, 1.5, 0.5, 1.2, 0.35, 2,0.7, 0.8, or 0.9 on a nominal available dose normalized basis; 2) 3.4 to9.2, 3.5 to 8.5, 5.5 to 9.2, 4.5 to 7.5, 5.8 to 7, 3.4, 3.5, 4.5, 5.5,9.2, 8.5, 7.5, 5.8, 5.9, 6, or 7 on a dose to lung normalized basis; 3)1.7 to 7.5, 3.2 to 4.1, 1.9 to 6, 3.2 to 7.5, 1.7 to 5.2, 3.6, 4.1, 3.2,1.9, 6, 3.2, 7.4, 7.5, 1.7, 5.2 or 5.3 on a dose to subject normalizedbasis; 4) 0.9 to 3.3, 1.7 to 2.2, 0,9 to 3, 1 to 3, 1.7 to 3.3, 1 to2.7, 1.9, 2.1, 2.2, 1.7, 0.9, 1, 2, 3, 2.9, 3, 2, 3.3, or 2.7 on anemitted dose normalized basis.

In some embodiments, on the basis of the non-normalized dose ofcorticosteroid, the Cmaxprovided by the corticosteroid solution is 1.6to 2, 1.5 to 3, 1.5 to 2.5, 1.5 to 2, 1.5, 1.6, 2, 2.5, or 3 fold higherthan the Cmaxprovided by the suspension-based formulation when the doseof corticosteroid in the solution and the suspension is approximatelythe same amount loaded. On the basis of normalization to the nominalavailable dose of corticosteroid, the Cmaxprovided by the corticosteroidsolution is 1.8 to 6.2, 1.5 to 6.5, 2 to 6.5, 1.5 to 5.5, 2 to 4, 1.5 to4, 1.5 to 3, 2.7, 3.3, 3.4, 1.5, 6.5, 2, 5.5, 4, or 3 fold higher thanthe Cmaxprovided by the suspension-based formulation. On the basis ofnormalization to the dose of corticosteroid to lung, the Cmaxprovided bythe corticosteroid solution is 1.4 to 4.3, 1.4 to 4.5, 1.5 to 4.5, 1.5to 3.5, 1.5 to 3, 1.4 to 3, 1.5 to 2.5, 1.5 to 2, 2, 2.3, 1.4, 4.5, 3.5,3, 1.5 or 2.5 fold higher than the Cmaxprovided by the suspension-basedformulation. On the basis of normalization to the dose of corticosteroidto subject, the Cmaxprovided by the corticosteroid solution is 2 to 3.5,2 to 5, 1.7 to 3.8, 1.7 to 5, 2.7, 3, 2.4, 2, 3.5, 5, 1.7, or 3.8 foldhigher than the Cmaxprovided by the suspension-based formulation. On thebasis of normalization to the emitted dose of corticosteroid, theCmaxprovided by the corticosteroid solution is 1.9 to 6.3, 1.75 to 6.5,2.2 to 4.2, 2.2 to 6.3, 1.9 to 4.2, 3.2, 3.5, 3.6, 2.8, 1.75, 6.5, 2.2,4.2, or 6.3 fold higher than the Cmaxprovided by the suspension-basedformulation.

In some embodiments, the Cmaxprovided by the corticosteroid solution isat least 1.5, 1.6, 2, 2.6, and 3 fold higher than the Cmaxprovided bythe suspension-based formulation when the dose of corticosteroid in thesolution is about 2 fold lower than the dose in the suspension.

In some embodiments, the formulation and system of the invention canprovide an AUC_(inf) (pg*h/ml) of 500 to 1700, 530 to 1650, 250 to 2500,280 to 1300, 780 to 1300, 980 to 2450, 275, 775, 980, 2400, 2500, 1300,1290, 530, 1650, 250, 280 or 780 on a dose non-normalized basis. Theformulation and system of the invention can provide a dose normalizedAUC_(inf) (pg/ml/μg) of: 1) 1 to 5.5, 2 to 2.2, 1 to 5.3, 1.1 to 5.2,1.5 to 2.6, 1.3 to 3.3, 2, 2.1, 2.2, 1, 5.5, 5.3, 5.2, 1.5, 2.6, 1.3 or3.3 on a nominal available dose normalized basis; 2) 10 to 25, 14 to 18,10.2 to 20, 13.6 to 18.8, 11.2 to 24.7, 10.2, 20, 13.6, 14, 19, 18.8,11, 11.2, 25, 24.7, 14.2, 16.2, 17.3 on a dose to lung normalized basis;3) 4 to 16, 4.2 to 16.1, 8 to 12.2, 5.4 to 16, 5.4 to 17, 8.5 to 9.6,8.5, 9.5, 9.6, 4.2, 16.1, 8, 12.2, 12, 5.4, 16, 17, or 16.5 on a dose tosubject normalized basis; 4) 2.5 to 9, 2.6 to 8.5, 4.5 to 5.1, 2.5 to 8,2.6 to 7.9, 4.2 to 6.7, 3.1 to 8.5, 3.2 to 8.5, 4.5, 4.6, 5, 5.1, 2.5,2.6, 4.2, 3.1, 9, 8.5, 5.1, 8, 7.9, or 6.7 to on an emitted dosenormalized basis.

In some embodiments, on the basis of the non-normalized dose ofcorticosteroid, the AUC_(inf) provided by the corticosteroid solution is1.6 to 2.5, 1.6 to 3.1, 1.5 to 3.5, 1.5 to 3.3, 2.5 to 3.3, 3.1, 1.5,3.3, 1.6, or 2.5 fold higher than the AUC_(inf) provided by thesuspension-based formulation when the dose of corticosteroid in thesolution and the suspension is approximately the same. On the basis ofnormalization to the nominal available dose of corticosteroid, theAUC_(inf) provided by the corticosteroid solution is 1.75 to 6.5, 1.75to 6.1, 2 to 6.5, 2 to 6.1, 2 to 4.5, 2 to 4.4, 3.3, 3.2, 3.5, 3.4,1.75, 6.5, 6.1, 2, 4.5, or 4.4 fold higher than the AUC_(inf) providedby the suspension-based formulation. On the basis of normalization tothe dose of corticosteroid to lung, the AUC_(inf) provided by thecorticosteroid solution is 1.2 to 3.5, 1.2 to 4, 1.2 to 3, 1.2 to 2.85,1.5 to 3.5, 1.4 to 3.5, 2, 2.2, 2.3, 2.4, 1.2, 3, 4, 2.85, 1.5, 3.5, or1.4 fold higher than the AUC_(inf) provided by the suspension-basedformulation. On the basis of normalization to the dose of corticosteroidto subject, the AUC_(inf) provided by the corticosteroid solution is 1.6to 4.9, 1.5 to 5, 1.6 to 5, 1.6 to 3.7, 1.6 to 3.6, 2 to 4.9, 1.9 to 4,2.6, 1.5, 5, 1.6, 3.7, 3.6, 2, 4.9, 1.9, or 4 fold higher than theAUC_(inf) provided by the suspension-based formulation. On the basis ofnormalization to the emitted dose of corticosteroid, the AUC_(inf)provided by the corticosteroid solution is 1.5 to 6, 1.7 to 6, 1.9 to 6,1.9 to 5.4, 2.3 to 5.8, 1.9 to 5.5, 1.9 to 5.8, 1.5, 6, 1.7, 1.9, 5.4,2.3, 5.8, 5.8, 3.2, 3.5, or 3.6 fold higher than the AUC_(inf) providedby the suspension-based formulation.

In some embodiments, the AUC_(inf) provided by the corticosteroidsolution is at least 1.5, 1.6, 2, 2.5, 3 and 3.1 fold higher than theAUC_(inf) provided by the suspension-based formulation when the dose ofcorticosteroid in the solution is about 2 fold lower than the dose inthe suspension.

The formulation and system of the invention can provide an AUC_(0-8 hr)(pg*h/ml) of 2000 to 3000, 2500 to 2700, 2000, 3000, or 2600 on a dosenon-normalized basis for the corticosteroid. The formulation and systemof the invention can provide a dose normalized AUC_(inf) (pg/ml/μg)of: 1) 2 to 3, 2.5 to 2.7, or 2.6 on a loaded dose nominal normalizedbasis for the corticosteroid; 2) 3 to 4, 3.4 to 3.5, or 3.4 on anemitted dose normalized basis for the corticosteroid.

In some embodiments, on the basis of the normalized nominal (loaded)dose of corticosteroid, the AUC₀₋₈ hr provided by the corticosteroidsolution is at least 1.7, 1.8, 1.9, or 2 fold higher than theAUC_(0-8 hr) provided by the suspension-based formulation when the doseof corticosteroid in the solution and the suspension is approximatelythe same. On the basis of normalization to the emitted dose ofcorticosteroid, the AUC_(0-8 hr) provided by the corticosteroid solutionis at least 1.5, 1.6, to 2 fold higher than the AUC_(0-8 hr) provided bythe suspension-based formulation.

The formulation and system of the invention can provide a C_(max)(pg/ml) of 1600 to 1800, 1650 to 1750, or 1700 on a dose non-normalizedbasis. The formulation and system of the invention can provide a dosenormalized C_(max) (pg/ml/μg) of: 1) 1 to 2, 1.6 to 1.8, or 1.7 on aloaded dose nominal normalized basis; 2) 2 to 2.5, or 2.2 on an emitteddose normalized basis.

In some embodiments, on the basis of the normalized nominal (loaded)dose of corticosteroid, the C_(max) provided by the corticosteroidsolution is at least 1.7, 1.8, 1.9, or 2 fold higher than the C_(max)provided by the suspension-based formulation when the dose ofcorticosteroid in the solution and the suspension is approximately thesame. On the basis of normalization to the emitted dose ofcorticosteroid, the C_(max) provided by the corticosteroid solution isat least 1.5, 1.6, to 2 fold higher than the C_(max) provided by thesuspension-based formulation.

In some embodiments, greater than 20%, greater than 25%, greater than29%, greater than 35% of the emitted dose may be absorbed into thebloodstream of the patient. In some embodiments, greater than 40%,greater than 50%, greater than 55% of the dose to subject may beabsorbed into the bloodstream of the patient. In some embodiments,greater than 10%, greater than 12%, greater than 15% of the nominalavailable dose may be absorbed into the bloodstream of the patient.

Some aspects of the invention provide a method of administering ordelivering a dose of corticosteroid to the air passageways of a subjectin need thereof, the method comprising administering or delivering thecorticosteroid with a nebulizer comprising a charge of an aqueous liquidformulation, the formulation comprising an aqueous liquid carrier,sulfoalkyl ether cyclodextrin, and the corticosteroid dissolved therein,wherein, during operation, the system provides enhanced drug delivery,increased rate of drug administration, reduced treatment time, reducedtoxicity, improved stability, enhanced bioabsorption, increased outputrate, increased total output, enhanced pharmacokinetic profile, reducedcorticosteroid-related side effects, enhanced pulmonary deposition,reduced oropharyngeal deposition, and/or improved nebulizationperformance over another system comprising the nebulizer and asuspension based formulation of the corticosteroid. The systems can beoperated under similar conditions or dissimilar conditions.

The invention includes all combinations of the embodiments and aspectsdisclosed herein. Accordingly, the invention includes the embodimentsand aspects specifically disclosed, broadly disclosed, or narrowlydisclosed herein, as well as combinations thereof and subcombinations ofthe individual elements of said embodiments and aspects.

These and other aspects of this invention will be apparent uponreference to the following detailed description, examples, claims andattached figures.

BRIEF DESCRIPTION OF THE FIGURES

The following drawings are given by way of illustration only, and thusare not intended to limit the scope of the present invention.

FIG. 1 depicts a graph of the hemolytic behavior of the CAPTISOL® ascompared to the same for the parent β-cyclodextrin, the commerciallyavailable hydroxypropyl derivatives, ENCAPSIN™ (degree of substitution˜3-4) and MOLECUSOL® (degree of substitution ˜7-8), and two othersulfobutyl ether derivatives, SBE1-β-CD and SBE4-β-CD.

FIG. 2 depicts a graph of the osmolality of SBE-CD containing solutionsof various degrees of substitution and HP-β-CD containing solutionscomprising similar concentrations of cyclodextrin derivative.

FIG. 3 depicts a phase solubility graph of the concentration (molar) ofcyclodextrin versus the concentration (molar) of budesonide for γ-CD,HP-β-CD and SBE7-β-CD.

FIG. 4 depicts a chart of the estimated percentage of nebulizationcomposition emitted from three different nebulizers (PARI LC PLUS,HUDSON UPDRAFT II NEB-U-MIST, and MYSTIQUE) for each of four differentnebulization compositions (PULMICORT RESPULES suspension, 5% w/vSBE7-β-CD solution, 10% w/v SBE7-β-CD solution and 20% w/v SBE7-β-CDsolution).

FIGS. 5 a-5 b depict droplet size data for nebulization of solutionswith a PARI LC PLUS nebulizer.

FIG. 6 depicts droplet size data for nebulization of solutions with aHUDSON UPDRAFT II NEBUMIST nebulizer.

FIG. 7 depicts droplet size data for nebulization of solutions with aMYSTIQUE ultrasonic nebulizer.

FIG. 8 depicts comparative Dv₅₀ droplet size data for nebulization ofcomposition with the three nebulizers PARI LC PLUS, HUDSON UPDRAFT IINEBUMIST, and MYSTIQUE.

FIG. 9 is a graph depicting the relationship between concentration ofSAE-CD versus output rate of SAE-CD in various different nebulizers.

FIGS. 10 a-10 b depict comparative droplet size data for nebulizationsolutions with the PARI LC PLUS and MYSTIQUE nebulizers of PULMICORTRESPULES suspension and a modified PULMICORT RESPULES-based SAE-CDsolution.

FIG. 11 depicts a semi-log plot of the % of initial concentration of theR- and S-isomers of budesonide in solutions with and without CAPTISOLversus time at 60° C. in solution.

FIG. 12 depicts a semi-log plot of the % of initial concentration ofbudesonide versus Lux hours when the samples are exposed to fluorescentlamps.

FIG. 13 depicts a phase solubility diagram for fluticasone propionate inthe presence of several different cyclodextrins.

FIG. 14 depicts a phase solubility diagram for mometasone furoate in thepresence of several different cyclodextrins.

FIG. 15 depicts a phase solubility diagram for esterified andnon-esterified fluticasone in the presence of SAE(5-6)-γ-CD.

FIG. 16 depicts a bar chart summarizing the aqueous solubility ofbeclomethasone dipropionate in the presence of various SAE-CDderivatives.

FIG. 17 depicts a plot of the geometric mean of dose (of corticosteroid,μg) to subject versus the geometric mean of AUC (pg*hr/ml) whenbudesonide is administered to subjects according to Example 17.

FIG. 18 depicts a plot of the geometric mean of dose (of corticosteroid,μg) to lung of subject versus the geometric mean of AUC (pg*hr/ml) whenbudesonide is administered to subjects according to Example 17.

FIG. 19 depicts a plot of dose (of corticosteroid, μg) to subject versusthe AUC (pg*hr/ml) for each individual when budesonide is administeredto subjects according to Example 17.

FIG. 20 depicts a plasma concentration profile for budesonide forindividual subjects according to Example 17.

FIG. 21 depicts a plasma concentration profile for budesonide for dogsaccording to Example 31.

FIG. 22 is a chart detailing the approximate distribution of a dose ofbudesonide upon administration of a solution of the invention accordingthe procedure of Example 17.

DETAILED DESCRIPTION OF THE INVENTION

The presently claimed formulation overcomes many of the undesiredproperties of other known aqueous inhalable solution or suspensioncorticosteroid-containing formulations. By including SAE-CD, in aninhalable liquid formulation containing corticosteroid, thecorticosteroid is dissolved. Unexpectedly, the nebulization ofcorticosteroid is improved in both an air driven jet nebulizer and anultrasonic nebulizer. Moreover, the corticosteroid exhibits greaterstability in the presence of SAE-CD than it does in its absence.

The inhalable formulation and system of the invention provides animproved pharmacokinetic profile over a suspension formulationcomprising approximately the same amount of corticosteroid and deliveredunder substantially the same conditions. The term “enhancedpharmacokinetic profile” is taken to mean a higher AUC (e.g. AUC_(last)or AUC^((0→∞)) per μg of corticosteroid delivered or administered, ahigher Cmaxper μg of corticosteroid delivered or administered, increasedbioavailability, absorption or distribution of the corticosteroid at thesite of delivery, a shorter Tmax or a longer Tmax.

Alternatively, the inhalable formulation and system of the inventionprovides substantially the same pharmacokinetic profile or an enhancedpharmacokinetic profile over a suspension formulation comprisingapproximately a higher amount of corticosteroid and delivered undersubstantially the same conditions. The corticosteroid in the formulationcan be present at a dose that is less than 80%, less than 70% less than60% less than 50%, less than 40%, less than 20%, and less than 10% ofthat in the suspension.

The corticosteroid can be present in an amount sufficient for singledose or multi-dose administration. SAE-CD can be present in an amountsufficient to solubilize the corticosteroid when the two are placed inthe aqueous carrier. The aqueous carrier can be present in an amountsufficient to aid in dissolution of the corticosteroid and form anebulization solution of sufficient volume and sufficiently lowviscosity to permit single dose or multi-dose administration with anebulizer. SAE-CD can be present in solid form or in solution in theaqueous carrier. The corticosteroid can be present in drypowder/particle form or in suspension in the aqueous carrier.

Commercially available air driven jet, ultrasonic or pulsating membranenebulizers include the AERONEB™ (Aerogen, San Francisco, Calif.),AERONEB GO (Aerogen); PARI LC PLUS™, PARI BOY™ N, PARI eflow, PARI LCSINUS, PARI SINUSTAR™, PARI SINUNEB, and PARI DURANEB™ (PARI RespiratoryEquipment, Inc., Monterey, Calif.); MICROAIR™ (Omron Healthcare, Inc,Vernon Hills, Ill.), HALOLITE™ (Profile Therapeutics Inc, Boston,Mass.), RESPIMAT™ (Boehringer Ingelheim Ingelheim, Germany) AERODOSE™(Aerogen, Inc, Mountain View, Calif.), OMRON ELITE™ (Omron Healthcare,Inc, Vernon Hills, Ill.), OMRON MICROAIR™ (Omron Healthcare, Inc, VernonHills, Ill.), MABISMIST™ II (Mabis Healthcare, Inc, Lake Forest, Ill.),LUMISCOPE™ 6610, (The Lumiscope Company, Inc, East Brunswick, N.J.),AIRSEP MYSTIQUE™, (AirSep Corporation, Buffalo, N.Y.), ACORN-1 andACORN-II (Vital Signs, Inc, Totowa, N.J.), AQUATOWER™ (MedicalIndustries America, Adel, Iowa), AVA-NEB (Hudson Respiratory CareIncorporated, Temecula, Calif.), AEROCURRENT™ utilizing the AEROCELL™disposable cartridge (AerovectRx Corporation, Atlanta, Ga.), CIRRUS(Intersurgical Incorporated, Liverpool, N.Y.), DART (ProfessionalMedical Products, Greenwood, S.C.), DEVILBISS™ PULMO AIDE (DeVilbissCorp; Somerset, Pa.), DOWNDRAFT™ (Marquest, Englewood, Colo.), FAN JET(Marquest, Englewood, Colo.), MB-5 (Mefar, Bovezzo, Italy), MISTY NEB™(Baxter, Valencia, Calif.), SALTER 8900 (Salter Labs, Arvin, Calif.),SIDESTREAM™ (Medic-Aid, Sussex, UK), UPDRAFT-II™ (Hudson RespiratoryCare; Temecula, Calif.), WHISPER JET™ (Marquest Medical Products,Englewood, Colo.), AIOLOS™ (Aiolos Medicnnsk Teknik, Karlstad, Sweden),INSPIRON™ (Intertech Resources, Inc., Bannockburn, Ill.), OPTIMIST™(Unomedical Inc., McAllen, Tex.), PRODOMO™, SPIRA™ (Respiratory CareCenter, Hameenlinna, Finland), AERx™ Essence™ and Ultra™, (AradigmCorporation, Hayward, Calif.), SONIK™ LDI Nebulizer (Evit Labs,Sacramento, Calif.), and SWIRLER® Radioaerosol System (AMICI, Inc.,Spring City, Pa.). Nebulizers that nebulize liquid formulationscontaining no propellant are suitable for use with the compositionsprovided herein. Any of these and other known nebulizers can be used todeliver the formulation of the invention including but not limited tothe following: nebulizers available from Pari GmbH (Starnberg, Germany),DeVilbiss Healthcare (Heston, Middlesex, UK), Healthdyne, Vital Signs,Baxter, Allied Health Care, Invacare, Hudson, Omron, Bremed, AirSep,Luminscope, Medisana, Siemens, Aerogen, Mountain Medical, AerosolMedical Ltd. (Colchester, Essex, UK), AFP Medical (Rugby, Warwickshire,UK), Bard Ltd. (Sunderland, UK), Carri-Med. Ltd. (Dorking, UK), PlaemNuiva (Brescia, Italy), Henleys Medical Supplies (London, UK),intersurgical (Berkshire, UK), Lifecare Hospital Supplies (Leies, UK),Medic-Aid Ltd. (West Sussex, UK), Medix Ltd. (Essex, UK), SinclairMedical Ltd. (Surrey, UK), and many other companies. The AERx andRESPIMAT nebulizers are described by D. E. Geller (Respir. Care (2002),47 (12), 1392-1404), the entire disclosure of which is incorporated byreference.

Nebulizers for use herein include, but are not limited to, jetnebulizers (optionally sold with compressors), ultrasonic nebulizers,vibrating membrane, vibrating mesh nebulizers, vibrating platenebulizers, vibrating cone nebulizer, and others. Exemplary jetnebulizers for use herein include Pari LC plus/ProNeb, Pari LCplus/ProNeb Turbo, Pari LC Plus/Dura Neb 1000 & 2000 Pari LCplus/Walkhaler, Pari LC plus/Pari Master, Pari LC star, Omron CompAir XLPortable Nebulizer System (NE-C18 and JetAir Disposable nebulizer),Omron compare Elite Compressor Nebulizer System (NE-C21 and Elite AirReusable Nebulizer, Pari LC Plus or Pan LC Star nebulizer with PronebUltra compressor, Pulomo-aide, Pulmo-aide LT, Pulmo-aide traveler,Invacare Passport, Inspiration Healthdyne 626, Pulmo-Neb Traverler,DeVilbiss 646, Whisper Jet, Acorn II, Misty-Neb, Allied aerosol, SchucoHome Care, Lexan Plasic Pocet Neb, SideStream Hand Held Neb, Mobil Mist,Up-Draft, Up-Draft II, T Up-Draft, ISO-NEB, Ava-Neb, Micro Mist, andPulmoMate. Exemplary ultrasonic nebulizers for use herein includeMicroAir, UltraAir, Siemens Ultra Nebulizer 145, CompAir, Pulmosonic,Scout, 5003 Ultrasonic Neb, 5110 Ultrasonic Neb, 5004 Desk UltrasonicNebulizer, Mystique Ultrasonic, Lumiscope's Ultrasonic Nebulizer,Medisana Ultrasonic Nebulizer, Microstat Ultrasonic Nebulizer, andMabismist Hand Held Ultrasonic Nebulizer. Other nebulizers for useherein include 5000 Electromagnetic Neb, 5001 Electromagnetic Neb 5002Rotary Piston Neb, Lumineb I Piston Nebulizer 5500, Aeroneb PortableNebulizer System, Aerodose™ Inhaler, and AeroEclipse Breath ActuatedNebulizer. Exemplary vibrating membrane, mesh or plate nebulizers aredescribed by R. Dhand (Respiratory Care, (December 2002), 47(12), p.1406-1418), the entire disclosure of which is hereby incorporated byreference.

Other suitable nebulizers are described in U.S. Pat. No. 5,954,047, No.6,026,808, U.S. Pat. No. 6,095,141, and U.S. Pat. No. 6,527,151, theentire disclosures of which are hereby incorporated by reference.

The present invention provides SAE-CD based formulations, wherein theSAE-CD is a compound of the Formula 1:

wherein:

-   n is 4, 5 or 6;-   R₁, R₂, R₃, R₄, R₅, R₆, R₇, R₈ and R₉ are each, independently, —O—    or a —O—(C₂-C₆ alkylene)-SO₃ ⁻ group, wherein at least one of R₁ to    R₉ is independently a —O—(C₂-C₆ alkylene)-SO₃ ⁻ group, preferably a    —O—(CH₂)_(m)SO₃ ⁻ group, wherein m is 2 to 6, preferably 2 to 4,    (e.g. —OCH₂CH₂CH₂SO₃ ⁻ or —OCH₂CH₂CH₂CH₂SO₃); and-   S₁, S₂, S₃, S₄, S₅, S₆, S₇, S₈ and S₉ are each, independently, a    pharmaceutically acceptable cation which includes, for example, H⁺,    alkali metals (e.g. Li⁺, Na⁺, K⁺), alkaline earth metals (e.g.,    Ca⁺², Mg⁺²), ammonium ions and amine cations such as the cations of    (C₁-C₆)-alkylamines, piperidine, pyrazine, (C₁-C₆)-alkanolamine and    (C₄-C₈)-cycloalkanolamine.

Exemplary embodiments of the SAE-CD derivative of the invention includederivatives of the Formula II (SAEx-α-CD), wherein “x” ranges from 1 to18; of the Formula III (SAEy-β-CD), wherein“y” ranges from 1 to 21; andof the Formula IV (SAEz-γ-CD), wherein“z” ranges from 1 to 24 such as:

SAEx-α-CD SAEy-β-CD SAEz-γ-CD Name SEEx-α-CD SEEy-β-CD SEEz-γ-CDSulfoethyl ether CD SPEx-α-CD SPEy-β-CD SPEz-γ-CD Sulfopropyl ether CDSBEx-α-CD SBEy-β-CD SBEz-γ-CD Sulfobutyl ether CD SptEx-α-CD SPtEy-β-CDSPtEz-γ-CD Sulfopentyl ether CD SHEx-α-CD SHEy-β-CD SHEz-γ-CD Sulfohexylether CD

“SAE” represents a sulfoalkyl ether substituent bound to a cyclodextrin.The values “x”, “y” and “z” represent the average degree of substitutionas defined herein in terms of the number of sulfoalkyl ether groups perCD molecule.

The SAE-CD used is described in U.S. Pat. No. 5,376,645 and U.S. Pat.No. 5,134,127 to Stella et al, the entire disclosures of which arehereby incorporated by reference. U.S. Pat. No. 3,426,011 to Parmerteret al. discloses anionic cyclodextrin derivatives having sulfoalkylether substituents. Lammers et al. (Recl. Trav. Chim. Pays-Bas (1972),91(6), 733-742); Staerke (1971), 23(5), 167-171) and Qu et al. (J.Inclusion Phenom. Macro. Chem., (2002), 43, 213-221) disclose sulfoalkylether derivatized cyclodextrins. U.S. Pat. No. 6,153,746 to Shah et al.discloses a process for the preparation of sulfoalkyl ether cyclodextrinderivatives. An SAE-CD can be made according to the disclosures ofStella et al., Parmerter et al., Lammers et al. or Qu et al., and ifprocessed to remove the major portion (>50%) of the underivatized parentcyclodextrin, used according to the present invention. The SAE-CD cancontain from 0% to less than 50% wt. of underivatized parentcyclodextrin.

The terms “alkylene” and “alkyl,” as used herein (e.g., in the—O—(C₂-C₆alkylene)SO₃ ⁻ group or in the alkylamines), include linear,cyclic, and branched, saturated and unsaturated (i.e., containing onedouble bond) divalent alkylene groups and monovalent alkyl groups,respectively. The term “alkanol” in this text likewise includes bothlinear, cyclic and branched, saturated and unsaturated alkyl componentsof the alkanol groups, in which the hydroxyl groups may be situated atany position on the alkyl moiety. The term “cycloalkanol” includesunsubstituted or substituted (e.g., by methyl or ethyl)cyclic alcohols.

An embodiment of the present invention provides compositions containinga mixture of cyclodextrin derivatives, having the structure set out informula (I), where the composition overall contains on the average atleast 1 and up to 3n+6 alkylsulfonic acid moieties per cyclodextrinmolecule. The present invention also provides compositions containing asingle type of cyclodextrin derivative, or at least 50% of a single typeof cyclodextrin derivative. The invention also includes formulationscontaining cyclodextrin derivatives having a narrow or wide and high orlow degree of substitution. These combinations can be optimized asneeded to provide cyclodextrins having particular properties.

The present invention also provides compositions containing a mixture ofcyclodextrin derivatives wherein two or more different types ofcyclodextrin derivatives are included in the composition. By differenttypes, is meant cyclodextrins derivatized with different types offunctional groups e.g., hydroxyalkyl and sulfoalkyl. Each independentdifferent type can contain one or more functional groups, e.g. SBE-CDwhere the cyclodextrin ring has only sulfobutyl functional groups, andhydroxypropyl-ethyl-β-CD where the cyclodextrin ring has bothhydroxypropyl functional groups and ethyl functional groups. The amountof each type of cyclodextrin derivative present can be varied as desiredto provide a mixture having the desired properties.

Exemplary SAE-CD derivatives include SBE4-β-CD, SBE7-β-CD, SBE11-β-CD,SBE3.4-γ-CD, SBE4.2-γ-CD, SBE4.9-γ-CD, SBE5.2-γ-CD, SBE6.1-γ-CD,SBE7.5-γ-CD, SBE7.8-γ-CD and SBE5-γ-CD which correspond to SAE-CDderivatives of the formula wherein n=5, 5, 5 and 6; m is 4; and thereare on average 4, 7, 11 and 5 sulfoalkyl ether substituents present,respectively. These SAE-CD derivatives increase the solubility of poorlywater soluble active agents to varying degrees.

Since SAE-CD is a poly-anionic cyclodextrin, it can be provided indifferent salt forms. Suitable counterions include cationic organicatoms or molecules and cationic inorganic atoms or molecules. The SAE-CDcan include a single type of counterion or a mixture of differentcounterions. The properties of the SAE-CD can be modified by changingthe identity of the counterion present. For example, a first salt formof SAE-CD can have a greater corticosteroid stabilizing and/orsolubilizing power than a different second salt form of SAE-CD.Likewise, an SAE-CD having a first degree of substitution can have agreater corticosteroid stabilizing and/or solubilizing power than asecond SAE-CD having a different degree of substitution. The enhancedsolubilization of a corticosteroid by one SAE-CD versus another isdemonstrated by the data in the following tables which depict the molarsolubility for fluticasone propionate with different SAE-CDs at about0.03 to 0.12M concentrations such that the solubilizing power followedabout this rank order over this concentration range of SAE-CD:SBE5.2-γ-CD>SPE5.4-γ-CD>SBE6.1-γ-CD>SBE9.7-γ-CD>>SBE7-α-CD>SBE6.7-β-CD>SPE7-β-CD.For mometasone furoate, the solubilizing power followed about this rankorder over this concentration range of SAE-CD:SBE9.7-γ-CD>SBE6.1-γ-CD>SBE5.2-γ-CD>>SPE5.4-γ-CD>SBE7-α-CD>SBE6.7-β-CD>SPE7-β-CD.Differences were also observed for the binding of budesonide andtriamcinolone with specific embodiments of SAE-CD. According to theinvention, a SAE-γ-CD binds a corticosteroid better than a SAE-β-CDdoes. Also, a SAE-β-CD binds budesonide better than a SAE-α-CD does. Thedata is summarized in FIGS. 13-14.

[Fluticasone] [Mometasone] ×10⁵ M ×10⁵ M [Triamcinolone [CD] as non asnon [Budesonide] acetonide] -CD M propionate esterified furoateesterified ×10⁵ M ×10⁵ M H₂O NA 0.39 0.16 1.82 0.00 6.59 3.56 β 0.015M1.36 12.9 81.3 (SBE)_(6.7) 0.0465 5.41 126.4 16.4 121.7 254.8 457.0 β0.0950 7.99 215.9 31.1 226.1 428.1 1023.3 (SBE)_(2.4) 0.04 1.70 12.8 β0.08 2.46 (SPE)₇ 0.04 1.05 93.9 7.23 122.4 β 0.08 2.12 151.2 10.8 223.3241.6

Solubility of Selected Steroids Enhanced by Alpha-Cyclodextrins

[Fluticasone] [Mometasone] ×10⁵ M ×10⁵ M [Triamcinolone [CD] as non asnon [Budesonide] acetonide] -CD M propionate esterified furoateesterified ×10⁵ M ×10⁵ M H₂O NA 0.39 0.16 1.82 0.00 6.59 3.56 A 0.040.00 8.4 0.08 0.27 28.5 (SBE)₇ 0.04 8.37 30.1 55.0 348.1 α 0.08 11.435.5 116.9 597.9

Solubility of Selected Steroids Enhanced by Gamma-Cyclodextrins

[Fluticasone] [Mometasone] ×10⁵ M ×10⁵ M [Triamcinolone [CD] as non asnon [Budesonide] acetonide] -CD M propionate esterified furoateesterified ×10⁵ M ×10⁵ M H₂O NA 0.39 0.16 1.82 0.00 6.59 3.56 Γ 0.03573.5 14.1 2.71 10.1 197.8 0.1 22.1 82.2 65.8 0.09 4.1 138.6 (SBE)_(5.2)0.04 79.12 375.8 γ 0.1 215.3 1440.4 93.9 889.2 861.6 (SBE)_(6.1) 0.0451.82 575.6 41.5 841.1 306.6 1059.5 γ 0.08 120.8 949.0 92.9 1423.1 698.82386.1 (SBE)_(9.7) 0.04 54.5 γ 0.075 103.1 895.0 94.0 889.6 453.4(SPE)_(5.4) 0.04 71.7 759.5 28.7 400.9 γ 0.08 140.1 1387.8 51.3 1467.1774.2

The inventors have also discovered that SAE-γ-CD is particularlysuitable for use in complexing esterified and non-esterifiedcorticosteroids as compared to complexation of the same corticosteroidswith SAE-β-CD or SAE-α-CD. The table above also summarizes the phasesolubility data depicted in FIG. 15 for fluticasone and fluticasonepropionate with various different SAE-γ-CD species having a degree ofsubstitution in the range of 5-10.

The present inventors have discovered that SAE-γ-CD is also much moreeffective at binding with a particular regioisomer of esterifiedcorticosteroids than is SAE-β-CD or SAE-α-CD. The procedure set forth inExample 18 details the comparative evaluation of the binding of SAE-γ-CDand SAE-β-CD with a series of structurally related corticosteroidderivatives. The table below summarizes the results of a study comparingthe binding of SBEx-γ-CD, wherein x represents the average degree ofsubstitution, derivatives and SBE-β-CD derivative with different formsof beclmethasone.

Beclomethasone Beclomethasone Beclomethasone 17-mono- 21-mono-Beclomethasone dipropionate propionate propionate (unesterified) CD(μg/mL) (μg/mL) (μg/mL) (μg/mL) SBE_(3.4) 0.04M →336.8 0.04M →10621.60.04M →172.6 0.04M →11360.2 γ-CD SBE_(5.24) 0.04M → 267.0 0.04M →9500.80.04M →139.8 0.04M →10949.9 γ-CD SBE_(6.1) 0.04M →243.8 0.04M →11666.90.04M →153.8 0.04M →11007.0 γ-CD SBE_(7.5) 00.04M → 168.5 0.04M →8539.10.04M →122.4 0.04M →9635.2 γ-CD SBE_(6.7) 0.04M →60.4 0.04M →6799.60.04M → 50.6 0.04M → 6927.0 β-CD γ-CD 0.04M →105.8 0.04M →136.9 0.04M→9.4 0.04M →114.8

The survey study shows that in the presence of SBE(3.4) γ-CD (0.04M),all of the forms of beclomethasone were at or near their highestsolubilities. B17P, the active metabolite of BDP, has the highestsolubility of the esterified beclomethasone forms in any of thederivatized CDs. The results indicate that SBE-γ-CD complexes withbeclomethasone dipropionate better than Captisol or γ-CD alone. Of theSAE-CD derivatives evaluated, the optimal degree of substitution of theSBE γ-CD that provides the greatest enhancement in solubility of BDP isDS=3.4, and solubility decreases almost linearly as the degree ofsubstitution increases. This is true for both the 24 hr and 5 dayequilibration times. So in terms of BDP solubilization with SAE-CD:SBE(3.4)γ-CD>SBE(5.2)γ-CD>SBE(6.1)γ-CD>SBE(7.5)γ-CD>γ-CD>Captisol(SBE7-β-CD). The data is summarized in FIG. 16. Therefore, the presentinventors have discovered that SAE-γ-CD cyclodextrin derivatives areunexpectedly better at solubilizing corticosteroids than are SAE-β-CDderivatives. Moreover, the formulations based upon SAE-γ-CD are suitablefor use in inhalable formulations contrary to the disclosure of Worth eta (above), which suggests that SAE-CD derivatives are not.

By “complexed” is meant “being part of a clathrate or inclusion complexwith”, i.e., a complexed therapeutic agent is part of a clathrate orinclusion complex with a cyclodextrin derivative. By “major portion” ismeant at least about 50% by weight. Thus, a formulation according to thepresent invention may contain an active agent of which more than about50% by weight is complexed with a cyclodextrin. The actual percent ofactive agent that is complexed will vary according to the complexationequilibrium constant characterizing the complexation of a specificcyclodextrin to a specific active agent. The invention also includesembodiments wherein the active agent is not complexed with thecyclodextrin or wherein a minor portion of the active agent is complexedwith the derivatized cyclodextrin. It should be noted that an SAE-CD, orany other anionic derivatized cyclodextrin, can form one or more ionicbonds with a positively charged compound. This ionic association canoccur regardless of whether the positively charged compound is complexedwith the cyclodextrin either by inclusion in the cavity or formation ofa salt bridge.

The binding of a drug to the derivatized cyclodextrin can be improved byincluding an acid or base along with the drug and cyclodextrin. Forexample, the binding of a basic drug with the cyclodextrin might beimproved by including an acid along with the basic drug andcyclodextrin. Likewise, the binding of an acidic drag with thecyclodextrin might be improved by including a base (alkaline material)along with the acidic drug and cyclodextrin. The binding of a neutraldrug might be improved by including a basic, acidic or other neutralcompound along with the neutral drug and cyclodextrin. Suitable acidiccompounds include inorganic and organic acids. Examples of inorganicacids are mineral acids, such as hydrochloric and hydrobromic acid.Other suitable acids include sulfuric acid, sulfonic acid, sulfenicacid, and phosphoric acid. Examples of organic acids are aliphaticcarboxylic acids, such as acetic acid, ascorbic acid, carbonic acid,citric acid, butyric acid, fumaric acid, glutaric acid, glycolic acid,α-ketoglutaric acid, lactic acid, malic acid, mevalonic acid, maleicacid, malonic acid, oxalic acid, pimelic acid, propionic acid, succinicacid, tartaric acid, or tartronic acid. Aliphatic carboxylic acidsbearing one or more oxygenated substituents in the aliphatic chain arealso useful. A combination of acids can be used.

Suitable basic compounds include inorganic and organic bases. Suitableinorganic bases include ammonia, metal oxide and metal hydroxide.Suitable organic bases include primary amine, secondary amine, tertiaryamine, imidazole, triazole, tetrazole, pyrazole, indole, diethanolamine,triethanolamine, diethylamine, methylamine, tromethamine (TRIS),aromatic amine, unsaturated amine, primary thiol, and secondary thiol. Acombination of bases can be used.

An anionic derivatized cyclodextrin can complex or otherwise bind withan acid-ionizable agent. As used herein, the term acid-ionizable agentis taken to mean any compound that becomes or is ionized in the presenceof an acid. An acid-ionizable agent comprises at least oneacid-ionizable functional group that becomes ionized when exposed toacid or when placed in an acidic medium. Exemplary acid-ionizablefunctional groups include a primary amine, secondary amine, tertiaryamine, quaternary amine, aromatic amine, unsaturated amine, primarythiol, secondary thiol, sulfonium, hydroxyl, enol and others known tothose of ordinary skill in the chemical arts.

The degree to which an acid-ionizable agent is bound by non-covalentionic binding versus inclusion complexation formation can be determinedspectrophotometrically using methods such as ¹HNMR, ¹³CNMR, or circulardichroism, for example, and by analysis of the phase solubility data forthe acid-ionizable agent and anionic derivatized cyclodextrin. Theartisan of ordinary skill in the art will be able to use theseconventional methods to approximate the amount of each type of bindingthat is occurring in solution to determine whether or not bindingbetween the species is occurring predominantly by non-covalent ionicbinding or inclusion complex formation. An acid-ionizable agent thatbinds to derivatized cyclodextrin by both means will generally exhibit abi-phasic phase solubility curve. Under conditions where non-covalentionic bonding predominates over inclusion complex formation, the amountof inclusion complex formation, measured by NMR or circular dichroism,will be reduced even though the phase solubility data indicatessignificant binding between the species under those conditions;moreover, the intrinsic solubility of the acid-ionizable agent, asdetermined from the phase solubility data, will generally be higher thanexpected under those conditions.

As used herein, the term non-covalent ionic bond refers to a bond formedbetween an anionic species and a cationic species. The bond isnon-covalent such that the two species together form a salt or ion pair.An anionic derivatized cyclodextrin provides the anionic species of theion pair and the acid-ionizable agent provides the cationic species ofthe ion pair. Since an anionic derivatized cyclodextrin is multi-valent,an SAE-CD can form an ion pair with one or more acid-ionizable agents.

The parent cyclodextrins have limited water solubility as compared toSAE-CD and HPCD. Underivatized α-CD has a water solubility of about14.5% w/v at saturation. Underivatized β-CD has a water solubility ofabout 1.85% w/v at saturation. Underivatized γ-CD has a water solubilityof about 23.2% w/v at saturation. Dimethyl-beta-cyclodextrin (DMCD)forms a 43% w/w aqueous solution at saturation. The SAE-CD can becombined with one or more other cyclodextrins or cyclodextrinderivatives in the inhalable solution to solubilize the corticosteroid.

Other water soluble cyclodextrin derivatives that can be used accordingto the invention include the hydroxyethyl, hydroxypropyl (including 2-and 3-hydroxypropyl) and dihydroxypropyl ethers, their correspondingmixed ethers and further mixed ethers with methyl or ethyl groups, suchas methylhydroxyethyl, ethyl-hydroxyethyl and ethyl-hydroxypropyl ethersof alpha-, beta- and gamma-cyclodextrin; and the maltosyl, glucosyl andmaltotriosyl derivatives of alpha, beta- and gamma-cyclodextrin, whichmay contain one or more sugar residues, e.g. glucosyl or diglucosyl,maltosyl or dimaltosyl, as well as various mixtures thereof, e.g. amixture of maltosyl and dimaltosyl derivatives. Specific cyclodextrinderivatives for use herein include hydroxypropyl-beta-cyclodextrin,hydroxyethyl-beta-cyclodextrin, hydroxypropyl-gamma-cyclodextrin,hydroxyethyl-gamma-cyclodextrin, dihydroxypropyl-beta-cyclodextrin,glucosyl-alpha-cyclodextrin, glucosyl-beta-cyclodextrin,diglucosyl-beta-cyclodextrin, maltosyl-alpha-cyclodextrin,maltosyl-beta-cyclodextrin, maltosyl-gamma-cyclodextrin,maltotriosyl-beta-cyclodextrin, maltotriosyl-gamma-cyclodextrin anddimaltosyl-beta-cyclodextrin, and mixtures thereof such asmaltosyl-beta-cyclodextrin/dimaltosyl-beta-cyclodextrin, as well asmethyl-beta-cyclodextrin. Procedures for preparing such cyclodextrinderivatives are well-known, for example, from Bodor U.S. Pat. No.5,024,998 dated Jun. 18, 1991, and references cited therein. Othercyclodextrins suitable for use in the present invention include thecarboxyalkyl thioether derivatives such as ORG 26054 and ORG 25969 madeby ORGANON (AKZO-NOBEL), hydroxybutenyl ether derivatives made byEASTMAN, sulfoalkyl-hydroxyalkyl ether derivatives, sulfoalkyl-alkylether derivatives, and other derivatives as described in US FragrantPatent Application Publications No. 2002/0128468, No. 2004/0106575, No.2004/0109888, and No. 2004/0063663, or U.S. Pat. No. 6,610,671, U.S.Pat. No. 6,479,467, U.S. Pat. No. 6,660,804, or U.S. Pat. No. 6,509,323.

The HP-β-CD can be obtained from Research Diagnostics Inc. (Flanders,N.J.). HP-β-CD is available with different degrees of substitution.Exemplary products include ENCAPSIN™ (degree of substitution˜4;HP4-β-CD) and MOLECUSOL™ (degree of substitution˜8; HP8-β-CD); however,embodiments including other degrees of substitution are also available.Since HPCD is non-ionic, it is not available in salt form.

Dimethyl cyclodextrin is available from FLUKA Chemie (Buchs, CH) orWacker (Iowa). Other derivatized cyclodextrins suitable in the inventioninclude water soluble derivatized cyclodextrins; Exemplary water-solublederivatized cyclodextrins include carboxylated derivatives; sulfatedderivatives; alkylated derivatives; hydroxyalkylated derivatives;methylated derivatives; and carboxy-β-cyclodextrins, e.g.succinyl-β-cyclodextrin (SCD), and6^(A)-amino-6^(A)-deoxy-N-(3-carboxypropyl)-β-cyclodextrin. All of thesematerials can be made according to methods known in the prior art.Suitable derivatized cyclodextrins are disclosed in ModifiedCyclodextrins: Scaffolds and Templates for Supramolecular Chemistry(Eds. Christopher J. Easton, Stephen F. Lincoln, Imperial College Press,London, UK, 1999) and New Trends in Cyclodextrins and Derivatives (Ed.Dominique Duchene, Editions de Santé, Paris, France, 1991).

Sulfobutyl ether β-cyclodextrin (CAPTISOL, CyDex Inc., degree ofsubstitution=6.6), 2-hydroxypropyl β-cyclodextrin (HP-β-CD, CERESTAR,degree of substitution=5.5), succinylated-β-cyclodextrin (S-CD,Cyclolab), and 2,6,di-o-methyl-β-cyclodextrin (DM-CD, Fluka) % w/wsolutions were prepared at their native pH or buffered as needed.Sulfoalkyl ether γ-CD and sulfoalkyl ether α-CD derivatives wereobtained from CyDex, Inc. (Lenexa, Kans.) and The University of Kansas(Lawrence, Kans.).

The amount of derivatized cyclodextrin required to provide the desiredeffect will vary according to the materials comprising the formulation.

Different cyclodextrins are able to solubilize a corticosteroid todifferent extents. FIG. 3 depicts a molar phase solubility curve forbudesonide with HP-β-CD, SBE7-β-CD, and γ-CD as compared to water. Theinventors have found that SAE-CD is superior to other cyclodextrins andcyclodextrin derivatives at solubilizing budesonide. On a molar basis,SBE-β-CD is a better solubilizer of budesonide than HP-β-CD. Inaddition, the solubilizing power among the SAE-CD derivatives followedabout this rank order for budesonide over a SAE-CD concentration rangeof 0.04 to 0.1M: SBE5.2-γ-CD˜SPE5.4-γ-CD>SBE6.1-γ-CD>SBE7-α-CD>SBE9.7-γ-CD SBE6.7-β-CD>SPE7-β-CD.For example, a 0.1M concentration of SBE7-β-CD was able to solubilize agreater amount of budesonide than either γ-CD or HP-β-CD. Moreover,SAE-CD-containing nebulizable formulations provide a greater output ratefor corticosteroid by nebulization as compared to γ-CD or HP-β-CDadministered under otherwise similar conditions.

It was unexpectedly discovered that the nebulization of Captisolsolutions provides several advantages with respect to othercyclodextrins. The droplets leaving the nebulizer are of a moreadvantageous size and the Captisol solutions are nebulized faster thansimilar solutions of other Cyclodextrins. The table below shows that theaverage particle size (Dv50) of Captisol solutions is smaller than thatof HP-β-CD or γ-CD. More importantly, as seen in the table below, theDv90 shows that the other cyclodextrins had significant number of verylarge droplets. The data (Malvern particle size) was obtained for eachformulation as emitted from a PARI LC PLUS nebulizer equipped with aPARI PRONEB ULTRA air compressor. The smaller droplet size is favoredfor an inhalable composition as it permits deeper lung delivery ofactive agents such as a corticosteroid.

Formulation Dv10 (μm) Dv 50 (μm) Dv 90 (μm) 5% Captisol  1.9 ± 0.04 3.84± 0.08 10.52 ± 0.2  10% Captisol 1.82 ± 0.05 3.61 ± 0.25 11.18 ± 1.9220% Captisol 1.78 ± 0.04 3.12 ± 0.11 10.02 ± 0.23 5% Hydroxypropyl 1.89± 0.04 3.99 ± 0.13 14.89 ± 2.45 β-Cyclodextrin 10% Hydroxypropyl 1.95 ±0.03 4.62 ± 0.34  120.1 ± 172.67 β-Cyclodextrin 20% Hydroxypropyl 1.91 ±0.02 4.26 ± 0.16 13.77 ± 1.00 β-Cyclodextrin 5% γ-Cyclodextrin 1.94 ±0.05 3.99 ± 0.36  205.62 ± 222.10 10% γ-Cyclodextrin 2.03 ± 0.05 4.84 ±0.49 451.55 ± 25.92 20% γ-Cyclodextrin 1.96 ± 0.04 4.97 ± 0.12  286.46 ±235.13

This advantage is further shown in the output rate of these solutions.The table below shows that Captisol is emitted from the nebulizer fasterand also to a greater extent than the other cyclodextrins, thus theoutput rate of the nebulizer is greater when Captisol is nebulized.

Percent Sputter Output Rate Formulation Emitted Time (min) mg/min 5%Captisol 56.42 3.81 296 10% Captisol 55.13 3.84 287 20% Captisol 50.564.06 249 5% Hydroxypropyl 43.32 4.14 209 β-Cyclodextrin 10%Hydroxypropyl 46.22 4.27 216 β-Cyclodextrin 20% Hydroxypropyl 46.90 4.01234 β-Cyclodextrin 5% γ-Cyclodextrin 52.74 5.41 195 10% γ-Cyclodextrin53.75 4.98 216 20% γ-Cyclodextrin 51.91 4.81 216 Nebulization is stoppedwhen the sound changes (time to sputter) or visible particles are nolonger produced. Sputter occurs at substantial completion of delivery ofsolution in the reservoir of the nebulizer.

The advantage of Captisol was further demonstrated by preparingsolutions containing budesonide dissolved in various cyclodextrins andcomparing their performance in nebulization to the performance ofcommercial PULMICORT® RESPULES®, a commercially availablesuspension-based unit dose formulation. The suspension obtained fromseveral unit dose ampoules of PULMICORT™ was pooled to form a multi-usesuspension based unit dose formulation, and SAE-CD (specifically,CAPTISOL), HP-β- or γ-cyclodextrin powder was added to achieve a 0.25mg/ml budesonide solution concentration. These budesonide-containingsolutions contained 5% w/v Captisol (P5C), 1% w/v gamma-CD (P1γCD) and5% w/v hydroxypropyl-beta-cyclodextrin (P5HPβCD). Each was prepared atleast 30 minutes prior to all testing, All three formulations wereclear, colorless solutions. (Note: a 250 mg/mL solution of budesonidecannot be achieved in a 5% w/v solution of γ-cyclodextrin as it exhibits“B” type solubility behavior) A 2 ml aliquot of the suspension orsolution was placed in the same Pari LC Plus nebulizer setup and theamount of budesonide in the emitted droplets was determined bycollecting them onto a filter and measuring the budesonide using HPLC.As shown in the table below, the total output rate (μg budesonidecollected/time to sputter) for each suspension or solution.

Total Output Rate SD Sample ID (μg/min) (μg/min) Pulmicort 33.85 3.85Pulmicort + 5% Captisol 44.04 1.42 Pulmicort + 5% HP-β-CD 21.37 2.44Pulmicort + 1% γ-CD 40.36 5.73

The output rate is highest for the Captisol solution indicating that anequivalent amount of drug can be delivered in a shorter period of time.Under the conditions used, β-CD is unable to solubilize an equivalentamount of corticosteroid due to the limited solubility of β-CD in water.

The present invention can be used with other suspension-based aqueousformulations, which formulations may be adapted for nasal delivery orpulmonary delivery. Exemplary suspension-based aqueous formulationsinclude the UDB formulation (Sheffield Pharmaceuticals, Inc.),VANCENASE™ AQ (beclomethasone dipropionate aqueous suspension; ScheringCorporation, Kenilworth, N.J.), ATOMASE™ (beclomethasone dipropionateaqueous suspension; Douglas Pharmaceuticals Ltd., Aukland, Australia),BECONASE™ (beclomethasone dipropionate aqueous suspension; GlaxoWellcome, NASACORT AQ™ (triamcinolone acetonide nasal spray, AventisPharmaceuticals), TRI-NASAL™ (triamcinolone acetonide aqueoussuspension; Muro Pharmaceuticals Inc.) and AEROBID-M™, (flunisolideinhalation aerosol, Forest Pharmaceuticals), NASALIDE™ and NASAREL™(flunisolide nasal spray, Wax Corporation), FLONASE™ (fluticasonepropionate, GlaxoSmithKline), and NASONEX™ (mometasone furoate,Schering-Plough Corporation).

The suspension formulation can comprise corticosteroid present inparticulate, microparticulate, nanoparticulate or nanocrystallirie form.Accordingly, an SAE CD can be used to improve the administration of acorticosteroid suspension-based unit dose formulation. Moreover, theSAE-CD outperforms other cyclodextrin derivatives.

According to one embodiment, SAE-CD (in solid or liquid form) and asuspension-based unit dose formulation comprising corticosteroid aremixed. The SAE-CD is present in an amount sufficient to increase theamount of solubilized corticosteroid, i.e. decrease the amount ofunsolubilized corticosteroid, therein. Prior to administration, theliquid may be optionally aseptically filtered or terminally sterilized.The liquid is then administered to a subject by inhalation using anebulizer. As a result, the amount of drug that the subject receives ishigher than the subject would have received had the unaltered suspensionformulation been administered.

According to another embodiment, SAE-CD (in liquid form, as ready-to-useliquid or as a concentrate) and a solid unit dose formulation comprisingcorticosteroid are mixed to form a liquid formulation. The SAE-CD ispresent in an amount sufficient to solubilize a substantial portion ofthe corticosteroid. The liquid is then administered via inhalation usinga nebulizer.

According to another embodiment, SAE-CD (in solid form) and a solid unitdose formulation comprising corticosteroid are mixed to form a solidmixture to which is added an aqueous liquid carrier in an amountsufficient to form a nebulizable formulation. Mixing and/or heating areoptionally employed upon addition of the liquid carrier to form theformulation. The SAE-CD is present in an amount sufficient to solubilizea substantial portion of the corticosteroid. The formulation is thenadministered via inhalation using a nebulizer.

The size of the reservoir varies from one type of nebulizer to another.The volume of the liquid formulation may be adjusted as needed toprovide the required volume for loading into the reservoir of aparticular type or brand of nebulizer. The volume can be adjusted byadding additional liquid carrier or additional solution containingSAE-CD. In general, the reservoir volume of a nebulizer is about 10 μlto 100 ml. Low volume nebulizers, such as ultrasonic and vibratingmesh/vibrating plate/vibrating cone/vibrating membrane nebulizers,pre-filled reservoir strips inclusive of delivery nozzle typically havea reservoir volume of 10 μl to 6 ml or 10 μl to 5 ml. The low volumenebulizers provide the advantage of shorter administration times ascompared to large volume nebulizers.

Example 28 details a procedure for preparation of a solution of theinvention to be used with a low volume (low reservoir volume and/or lowreservoir residual volume) nebulizer, such as an AERx nebulizer. Thesolutions of the invention can be nebulized with any nebulizer; however,with an AERx delivery system that coordinates both the inhalation anddelivery processes to optimize deep lung penetration, an initial samplevolume of about 10 to 100, or 50 μl can be used to load AERx Strip unitdose container. Administration of this solution with the system makes itfeasible for a therapeutic dose to be administered to a subject in asingle puff (a single full inhalation by a subject, i.e. 3-5 seconds)via nebulization. Based on general performance expectations of suchdevices where the emitted dose is 60-65% and the subsequent lung dosesare 65-85%, about 10 μg to 60 μg or 10 μg to 110 μg of budesonide may beexpected to be delivered to the lung in a single dosing event usingbudesonide solutions prepared with SAECD.

Example 32 details a procedure for the comparison of nebulizationparameters in four different nebulizers using a formulation of theinvention and PULMICORT RESPULES (suspension-based formulation). In eachcase, the formulation of the invention out performs the suspension-basedformulation. The solution of the invention provide a 1.25, 1.4, 2.1,3.3, 3.67, 1.25 to 3.7, or 1.25 to 4 fold increase in the amount ofbudesonide delivered. Under the conditions tested, the AIRSEP MYSTIQUEwas most efficient at emitting/nebulizing the SAE-CD/budesonideformulation.

Unless otherwise specified, the term “dose”, which is understood toinclude an effective dose, is taken to mean a nominal dose, emitteddose, nominal available dose, dose to subject, dose to lung or othersuch term of art. The term “nominal dose” refers to an amount ofcorticosteroid placed in the reservoir of a nebulizer, wherein thevolume of liquid in the reservoir is determined according the size ofthe reservoir. The term “nominal available dose” refers to the amount ofcorticosteroid that is determined could be or should have been availableto a subject when administered a formulation of the invention bynebulization but formulation is/was not administered in its entirety.The term “emitted dose” refers to the amount of corticosteroid emittedfrom a nebulizer. The term “dose to subject” refers to the amount ofcorticosteroid delivered to and retained by a subject followingadministration of a formulation of the invention by nebulization. Theterm “dose to lung” refers to the amount of corticosteroid delivered toand retained by the lungs of a subject following administration of aformulation of the invention by nebulization.

In general, a single-use suspension-based unit dose formulation ofcorticosteroid contains about 0.125, 0.25, 0.5, 1, 2, or about 0.125 toabout 2 mg of corticosteroid suspended in about 50 μl to 10 ml of liquidcarrier. Alternatively, the corticosteroid is present at a concentrationof about 20 mcg to about 30 mg of corticosteroid per ml of suspension.As a result, about 10 to 500 mg of SAE-CD, or 10 to 250 mg of SAE-CD, or10 to 300 mg of SAE-CD, be it in solid form or dissolved in a liquidcarrier, is added to each ml the suspension in order to dissolve asubstantial portion of the corticosteroid and form a nebulizable unitdose liquid formulation that is then administered to a subject.

The formulation, method or system can employ a dose of about 1 μg to 20mg, 1 μg to 10 mg, 0.01 mg to 10 mg, 0.025 mg to 10 mg, 0.05 mg to 5 mg,0.1 mg to 5 mg, 0.125 mg to 5 mg, 0.25 mg to 5 mg, 0.5 mg to 5 mg, 0.05mg to 2 mg, 0.1 mg to 2 mg, 0.125 mg to 2 mg, 0.25 mg to 2 mg, 0.5 mg to2 mg of corticosteroid. These dose amounts are suitable for thecorticosteroids of the invention, in particular corticosteroids that areas lipophilic as or more lipophilic than flunisolide, whichcorticosteroids include beclomethasone, beclomethasone dipropionate,beclomethasone monopropionate, budesonide, ciclesonide,desisobutyryl-ciclesonide, flunisolide, fluticasone, fluticasonepropionate, mometasone, mometasone furoate, triamcinolone acetonide.

The formulation of the invention comprises a dose of corticosteroid inan approximate solution volume of 10 μl to 100 ml, 50 μl to 50 ml, 50 μlto 10 ml, 0.1 to 10 ml, 0.1 ml to less than 10 ml, 0.1 ml to 7.5 ml, 0.1ml to 5 ml, 0.1 ml to 3 ml, 0.1 ml to 2 ml, 0.1 ml to 1 ml, 0.05 ml to7.5 ml, 0.05 ml to 5 ml, 0.05 ml to 3 ml, 0.05 ml to 2 ml, or 0.05 ml to1 ml.

Due to the wide range of nebulizer reservoir volumes available and ofvarying dose requirements among the corticosteroids, a formulation ofthe invention can comprise 1 μg to 20 mg of corticosteroid in 0.01 ml to100 ml of solution volume.

In general, a multi-use suspension-based unit dose formulation ofcorticosteroid contains approximately 0.125 to 2 mg of corticosteroidsuspended in 1 to 100 ml of liquid carrier. A multi-use formulationactually contains two or more unit doses of corticosteroid. Single unitdose aliquots are taken from a multi-use unit dose formulation, and thesingle unit dose are typically administered one-at-a-time to a subject.As a result, about 10 to 500 mg of SAE-CD, be it in solid form ordissolved in a liquid carrier, is added to each ml the suspension inorder to dissolve a substantial portion of the corticosteroid and form amulti-use unit dose liquid formulation that is then administered to asubject in single unit dose aliquots.

One aspect of the invention is that a suspension-based unit doseformulation is converted to a liquid unit dose formulation prior topulmonary administration via inhalation (of a nebulized mist) to asubject. The conversion can take place in the same container in whichthe suspension is provided, in a different container, or in thereservoir of a nebulizer. In order to form a liquid formulation, asubstantial portion of the corticosteroid must be dissolved. As used inreference to the amount of dissolved corticosteroid, a “substantialportion” is at least 20% wt., at least 30% wt., at least 40% wt., or atleast 20% wt and less than 50% wt. of the corticosteroid. As used inreference to the amount of dissolved corticosteroid, a “major portion”is at least 50% wt. of the corticosteroid.

It is well known that pharmacists working in compounding pharmacies canand do prepare a suspension-based unit dose formulation comprisingcorticosteroid. Such pharmacists will now be able to prepare a singleuse or multi-use liquid unit dose formulation by employing a methoddescribed herein. Alternatively, a subject (patient) undergoingcorticosteroid treatment may convert the suspension-based formulation toa liquid formulation of the invention by employing a method describedherein. Instead of preparing the liquid formulation from the suspensionat the pharmacy, a kit containing the suspension formulation and SAE-CDcan be prepared.

The concentration of SAE-CD in solution can be expressed on a weight toweight or weight to volume basis; however, these two units areinterconvertible. When a known weight of cyclodextrin is dissolved in aknown weight of water, the % w/w cyclodextrin concentration isdetermined by dividing the cyclodextrin weight in grams by the totalweight (cyclodextrin+water weight) in like units and multiplying by 100.When a known weight of cyclodextrin is dissolved to a known totalvolume, the % w/v cyclodextrin concentration is determined by dividingthe cyclodextrin weight in grams by the total volume in milliliters andmultiplying by 100. The correlation between the two cyclodextrinconcentration percentages was experimentally determined by preparingvarious % w/w cyclodextrin solutions and measuring the density of eachwith a pycnometer at 25° C. The density (g/mL) of each % w/w CAPTISOLsolution is presented in the table below.

Captisol Density Viscosity % w/w (g/mL) (Cp, 25 C.) 59.4 1.320 527.049.4 1.259 51.9 39.7 1.202 17.0 29.8 1.149 5.91 19.7 1.095 2.78 8.51.041 1.75 0.0 1.002 1 slope = 0.0053 y-intercept = 0.995 correlation =0.9989

The resulting linear relationship readily enables the conversion ofCAPTISOL concentrations expressed in % w/w to that of % w/v by thefollowing equation:

% w/v=((% w/w*slope)+y-intercept)*% w/w

where the slope and intercept values are determined from a linearregression of the density data in the table. For example, by using theabove equation, a 40% w/w CAPTISOL solution would be equivalent to a˜48.3% w/v CAPTISOL solution.

The performance of an inhalable solution of the invention in a nebulizermay depend upon the viscosity of the solution in its reservoir, thenebulization solution. The viscosity of an aqueous solution of SBE7-β-CDchanges with respect to concentration approximately as indicated in thetable above. Viscosity of the inhalable composition can have an impacton percentage of nebulization composition emitted from a nebulizer,output rate of nebulized corticosteroid and droplet size distribution.

The amount of residual nebulization inhalable composition left in thereservoir of the nebulizer may be greater for solutions containingSAE-CD than for a budesonide-containing suspension. For example, FIG. 4depicts a chart of the estimated percentage of nebulization compositionemitted from three different nebulizers (PARI LC PLUS, HUDSON UPDRAFT IINEB-U-MIST, and MYSTIQUE) for each of four different nebulizationcompositions (PULMICORT RESPULES suspension, 5% w/w SBE7-β-CD solution,10% w/w SBE7-β-CD solution and 20% w/w SBE7-β-CD solution). ThePULMICORT RESPULES suspension was used as the control. The PARI LC PLUS,MYSTIQUE and HUDSON nebulizers were used for the comparison. TheMYSTIQUE nebulizer was unable to nebulize the suspension andconcentrated SAE-CD solution (20% w/w) efficiently so they were notevaluated with that nebulizer. The results suggest that, under theconditions tested, nebulization of PULMICORT RESPULES suspension resultsin a greater percentage of nebulized composition, meaning that, with thesuspension, less nebulization composition is left in the reservoir ofthe nebulizer upon completion of nebulization as compared to with thesolution. In some cases, nebulization of the suspension resulted in thegreatest percentage by weight of total composition emitted by anebulizer. In other words, under similar nebulization conditions, thePARI LC PLUS and HUDSON nebulizers more efficiently reduced the volumeof nebulization suspension than of nebulization solution; however, thisdid not correspond with the total amount of drug emitted by thenebulizer.

Output rate of an SAE-CD nebulization solution versus that of asuspension, each containing budesonide, was compared. A modified versionof the method of Example 10 was followed to determine output rate. Thetables below summarize the data observed.

2 minutes Total Bud Total Neb Total Neb Out Put Rate Total Out Budrecvr'd Recovered Time Time 2 minutes Put Rate Sample ID (ug) (ug)(min:sec) (min) (ug/min) (ug/min) 1-PUL-1 84.021 164.199 5:34 5.57 42.0129.48 1-PUL-2 90.395 175.63 4:58 4.97 45.20 35.34 1-PUL-3 82.046 174.5464:45 4.75 41.02 36.75 Mean 171.458 Mean 42.74 33.85 SD 6.310 SD 2.183.85 CV 3.680 CV 5.10 11.38 2-P5C-1 131.412 258.894 5:42 5.7 65.71 45.422-P5C-2 126.945 246.987 5:36 5.6 63.47 44.10 2-P5C-3 128.464 236.3715:33 5.55 64.23 42.59 Mean 247.417 Mean 64.47 44.04 SD 11.268 SD 1.141.42 CV 4.554 CV 1.76 3.22

Data obtained using a PARI LC PLUS nebulizer equipped with a PART PRONEBULTRA air compressor.

2 minutes Total Bud Total Neb Total Neb Out Put Rate Total Out Budrecvr'd Recovered Time Time 2 minutes Put Rate Sample ID (ug) (ug)(min:sec) (min) (ug/min) (ug/min) 10-PUL-1 11.200 27.926 5:20 5.33 5.605.24 10-PUL-2 29.015 40.11 4:15 4.25 14.51 9.44 10-PUL-3 25.363 30.5164:17 4.28 12.68 7.13 Mean 32.851 Mean 10.93 7.27 SD 6.419 SD 4.71 2.10CV 19.539 CV 43.05 28.93 11-P5C-1 41.049 98.155 5:47 5.78 20.52 16.9811-P5C-2 44.495 131.8 6:00 6 22.25 21.97 11-P5C-3 53.374 132.31 5:555.92 26.69 22.35 Mean 120.755 Mean 23.15 20.43 SD 19.574 SD 3.18 2.99 CV16.210 CV 13.73 14.66

Data obtained using a MYSTIQUE ultrasonic nebulizer.

All of the above formulations contain approximately 250 μg/mL, ofbudesonide. The samples identified as “P5C” contain 50 mg/mL (or about5%) SBE7-β-CD.

The table below shows the nebulizer output rate for solutions containingvarious levels of SAE-CD.

Nebulization Vis- Nebulizer % Emitted Time cosity Volume (By Weight(Minutes: Output Sample ID (Cp) (ml) Difference) Seconds) Rate 21.5% w/w3.06 3 47.47 10:51  4.52 SBE7-β-CD 10.75% w/w 1.84 3 51.36 8:53 6.02SBE7-β-CD 5.15% w/w 1.23 3 55.47 9:59 5.78 SBE7-β-CD H₂O 3 50.36 9:215.47

Surprisingly, nebulization of the SAE-CD-containing solution provided ahigher budesonide output rate than nebulization of the PULMICORTRESPULES suspension even though the nebulizer emitted a greater totalamount of the suspension. Higher SAE-CD concentrations, above 25% w/vled to slightly longer nebulization times and lower output rates oncethe viscosity exceeded an approximate upper limit.

Without being held bound to a particular mechanism, it is believed thatthe nebulizer preferentially nebulizes the water (supernatant) of thesuspension rather than the particles of the suspension thereby causingan increase in the molar concentration of budesonide in the suspensionin the reservoir. Since the corticosteroid is dissolved in the solutionof the invention, this type of separation does not occur to asignificant degree. As a result, the rate of increasing concentration ofcorticosteroid in the reservoir is lower for a solution than it is witha suspension. Further support for this phenomenon is detailed in thefollowing table.

RIC over RIC over 2 min. TTS, Initial Final mcg/mL mcg/mL Conc Conc permin per min Pulmicort 250 572 31.26 64.37 Respules CEB 250 398 14.9626.69

The data above was obtained by comparing the nebulization of a solutionof the invention (CEB, SAE-CD, 5% wt./v, 2 ml; budesonide, 250 μg/ml)and PULMICORT RESPULES suspension 2 ml; budesonide, 250 μg/ml) using aPARI LC+ equipped with a PARI PRONEB ULTRA air compressor. Theconcentration of budesonide in the reservoir was measured at two minutesand at time to sputter (TTS). The rate of increasing concentration (RIC)of the corticosteroid in the solution in the reservoir was thencalculated for each formulation over each interval of time. The RIC ofthe solution of the invention provided an approximately 48% decrease inthe RIC over two minutes and an approximately 42% decrease in the RICover the TTS. In some embodiments, the percentage decrease in the RICranges from 10% to 60%, 15% to 60%, 20% to 60%, 30% to 60%, or 40% to60%. In some embodiments, the system may provide a RIC of 0 to 40, 1 to40, 5 to 30, or 10 to 30 mcg of corticosteroid/mL of solution volume permin of operation/nebulization.

Accordingly, the invention provides a system for administration of acorticosteroid by nebulization comprising: a nebulizer comprising areservoir; and an inhalable aqueous composition comprising SAE-CD and adose of corticosteroid, whereby the system provides a reduced RIC forthe corticosteroid in the reservoir as compared to the RIC provided by asuspension comprising approximately the same amount of corticosteroidand volume of aqueous composition operated under approximately the sameconditions.

Based on data above, 21.5±5% w/w SBE7-β-CD concentration was identifiedas the approximate upper acceptable level for the nebulizer tested,“acceptable” being defined as the upper concentration of SBE7-β-CD thatcan be used without building up excessive viscosity, which may adverselyaffect the nebulization time and output rate. The practical upper limitfor concentration of SAE-CD will vary among the nebulizer formats. Theupper acceptable concentration of SAE-CD in a liquid formulation for usein a nebulizer may vary according to the DS of the derivative, the alkylchain length of the sulfoalkyl functional group, and/or the CD ring sizeof the SAE-CD.

For administration to the respiratory tract, particularly the lungs, anebulizer is used to produce appropriately sized droplets. Typically,the particle size of the droplet produced by a nebulizer for inhalationis in the range between about 0.5 to about 5 microns. If it is desiredthat the droplets reach the lower regions of the respiratory tract,i.e., the alveoli and terminal bronchi, the particle size range can bebetween about 0.5 and about 2.5 microns. If it is desired that thedroplets reach the upper respiratory tract, the particle size range canbe between 2.5 microns and 5 microns.

As noted above, viscosity of the nebulization composition, can impactdroplet size and droplet size distribution. For example, the presentformulations tend to form larger droplets, in terms of Dv50, at thelower concentrations, and thereby lower viscosity, of SAE-CD in theabsence of budesonide. FIGS. 5 a-5 b depict droplet size data fornebulization of inhalable compositions with a PARI LC PLUS nebulizer.For each of the figures, a MALVERN laser light scattering device(Mastersizer S, Malvern Instruments Ltd. Malvern, Worcs, U.K.) was usedto measure MMAD. FIG. 5 a depicts the results obtained using γ-CDsolutions of varying concentrations (5% w/v, 10% w/v and 20% w/v) in theabsence of budesonide. The results indicate that γ-CD on its own wouldnot behave acceptably in a nebulizer, since almost all of the mass ofthe solution is of an unacceptable droplet size range. Even withextensive recycling and droplet size selection by a nebulizer, a γ-CDbased nebulization solution containing corticosteroid would require anextremely long dosing period due to the low percentage of mass that isof the appropriate droplet size range, especially since γ-CD is not aneffective solubilizer of budesonide at the concentrations tested.

In comparison, FIG. 5 b depicts the results obtained using the samenebulizer with PULMICORT RESPULES suspension or a modified PULMICORTRESPULES solution containing SAE-CD of different concentrations (5% w/v,10% w/v and 20% w/v). With each of these samples, a significant portionof the nebulized mass is of a respirable size range. Moreover, thesolutions containing SAE-CD apparently form droplets that are comparablein size to those of the nebulized suspension.

FIG. 6 depicts droplet size data for nebulization of inhalablecompositions with a HUDSON UPDRAFT II NEBUMIST nebulizer charged withPULMICORT RESPULES suspension or a solution containing SAE-CD atdifferent concentrations. (5% w/v, 10% w/v and 20% w/v). As compared tothe PARI LC PLUS nebulizer, the NEB-U-MIST forms a slightly largerparticle size distribution, a significant portion of the nebulized massis still in the appropriate size range. Accordingly, the nebulizationsolution made from the suspension and containing SAE-CD is suitable foruse in a variety of different air driven jet nebulizers.

The package insert for PULMICORT RESPULES suspension states that thesuspension should not be nebulized with an ultrasonic nebulizer. FIG. 7depicts droplet size data for nebulization of inhalable compositionswith a MYSTIQUE ultrasonic nebulizer. The compositions include threedifferent SAE-CD containing solutions. Unlike the suspension, the SAE-CDcontaining solution can be nebulized with an ultrasonic nebulizer. Thus,the invention provides a method of improving the pulmonary delivery ofcorticosteroid in a suspension-based unit dose formulation from anultrasonic nebulizer, the method comprising the step of including SAE-CDin the formulation in an amount sufficient to decrease the amount ofundissolved corticosteroid in the suspension-based unit doseformulation.

The performance of nebulization compositions across a range ofnebulizers is typically compared by comparing the Dv50 of the dropletsize distribution for the respective compositions. FIG. 8 depictscomparative Dv50 droplet size data for nebulization of an inhalablecomposition with the three above-mentioned nebulizers. In each case, theSAE-CD containing solutions are suitable for administration bynebulization across a range of concentrations. Moreover, the dropletsize distribution can be partially controlled by adjusting theconcentration of SAE-CD.

FIG. 9 is a graph depicting the relationship between concentration ofSAE-CD versus output rate of SAE-CD in various different nebulizers withdifferent sources of compressed air required for the specific setup: theRAINDROP-Rat, RAINDROP-Dog, PARI LC STAR-UNC, PARI LC STAR-Rat PARI LCPLUS and DEVILBISS PULMO AIDE air jet driven nebulizers. The nebulizerswere used in a variety of setups including free standing as well asanimal exposure chambers and/or individual exposure masks. In general,the data demonstrate that output of SAE-CD increases with increasingSAE-CD concentration. Depending upon the nebulizer used, the conditionsunder which the nebulizer is operated and the concentration of SAE-CD insolution, different maximum output rates can be achieved. For example,the maximum output rate in the Raindrop-Dog setup is from a 250 mg/mLCAPTISOL concentration.

Even though nebulization of PULMICORT RESPULES suspension with anultrasonic nebulizer is not recommended, it can be achieved. FIGS. 10a-10 b depict comparative droplet size data for nebulization solutionswith the PARI LC PLUS and MYSTIQUE nebulizers of PULMICORT RESPULESsuspension and a modified PULMICORT RESPULES-based SAE-CD solution.PULMICORT RESPULES suspension with and without 5% w/v SBE7-β-CD wereused as the test samples. The procedure of Example 12 was followed. FIG.10 a depicts the Dv10 and Dv50 data for the solutions run on the PARI LCPLUS air driven jet nebulizer and FIG. 10 b depicts the Dv10 and Dv50data for the solutions run on the MYSTIQUE ultrasonic nebulizer. In eachcase, the droplet size data for the two different solutions iscomparable. However, the budesonide output rate for the two solutionswas significantly different. Use of SAE-CD in a nebulizationcomposition, however, results in an increased output rate of budesonideregardless of the format of the nebulizer. The invention, thus, providesa method of increasing the output rate of a corticosteroid-containingsuspension-based unit dose formulation being delivered by a nebulizer,the method comprising the step of including SAE-CD in the formulation inan amount sufficient to increase the amount of dissolved corticosteroidin the formulation to form an altered formulation, whereby the outputrate of corticosteroid for the altered formulation is greater than theoutput rate of corticosteroid for the suspension formulation.

Two pulsating membrane nebulizers (AERONEB GO and the nebulizer of U.S.Pat. No. 6,962,151) were evaluated according to Examples 21 and 22,respectively, to determine the performance of the devices with asolution of the invention and to demonstrate the utility of usingconcentrated solutions of corticosteroids in an efficient electronicnebulizer. Several in vitro parameters, important for the clinical useof corticosteroids, were determined and compared to these parameters ofthe solution formulation described in Example 6 and the 2 mL commercialreference suspension both in an air jet nebulizer. The totaldelivery/nebulization time (time to sputter where production of aerosolis no longer visible), MMAD droplet size and fine particle fractionparameters were obtained for each device. A cascade impactor was engagedwith the output of each device to determine performance duringnebulization and characterize the in vitro aerosol drug output.

The solution used in the AERONEB GO device was compared side-by-sidewith a RAINDROP air jet nebulizer employing a solution (0.5 ml)comprising budesonide (500 μg/0.5 ml), CAPTISOL (10% w/v), water, andTween 80 (optional), wherein the solution was made by adding CAPTISOL toa suspension of budesonide particles (RESPULES) as detailed below. Ineach device, the control suspension-based sample was the RESPULESsuspension (2 ml containing 500 mg of budesonide). The RAINDROPnebulizer was equipped with a PARI PRONEB ULTRA compressor. The AERONEBGO micropump nebulizer was equipped with the OnQ Aerosol Generator. Thetotal output of budesonide at cessation of aerolization (at sputter) wasquantified. The emitted dose and fine particle dose were measured withthe cascade impactor. The solution in the nebulizer of the '151 patentcomprised budesonide, CAPTISOL, water, and technetium-99 radiolabel inthe form of diethylenetriaminepenta-acetic acid as a surrogate markerfor budesonide. The data from the two studies is summarized in the tablebelow.

Aerosol Performance Using NGI Cascade Impactor Nebulizer device AeroNebGo Raindrop/Pari Proneb Ultra Formulation CEB #1 Pulmicort CEB #2Pulmicort Amount of drug (μg)* 500.00 500.00 500.00 500.00 NebulizationTime (min)+ 1.0 4.0 5.0 5.5 Amt of drug exiting 405.93 266.06 150.2792.42 nebulizer (μg)* Percent of drug exiting 81.26 53.24 30.03 18.49nebulizer (%) Amt of drug exiting nebulizer 262.17 131.73 131.44 76.27in fine particle fraction (% < 5 μm) (μg)* Percent of drug exiting 66.3349.54 87.43 82.55 nebulizer in fine particle fraction (<5 μm,) Percentof total drug in fine 53.91 26.38 26.27 15.26 particle fraction (< 5 μm,%) MMAD (um, estimated by 4.10 5.54 2.22 2.86 linear regression) GSD(sigma g, estimated by 2.02 1.55 2.37 2.21 linear regression) *Datanormalized to 500 μg budesonide in each nebulizer using total ofrecovered drug. +Note: Nebulizers were run 30 seconds past end ofnebulization time to insure complete emptying into the NGI impactor.

Using the electronic pulsating membrane nebulizers (AERONEB GO nebulizerand the nebulizer of the '151 patent), delivery of a unit dose ofcorticosteroid was completed within less than one minute. The RAINDROPnebulizer completed delivery of a unit dose in just over 5.5 min withthe RESPULES and just over 5 min with a solution of the invention. Inaddition, the pulsating membrane nebulizers delivered a substantiallygreater percentage of corticosteroid in the FPF, which is generallydefined as the fraction of particles less than 5μ a or the fraction ofparticles on cascade impactor stages with a cut-off of less than 6μ.Accordingly, the total nebulization time of the AERONEB GO is one fourththe time to sputter for the Pari LC+ air jet nebulizer. As a result,treatment time would be reduced with the pulsating membrane nebulizer ascompared to the air jet nebulizer, and the amount of budesonide emittedfrom the pulsating membrane nebulizer is 2 to 3 times more than from theair jet nebulizer. It was also determined that the percent of drugexiting the nebulizer (the emitted dose) was 81% of the amount initiallyloaded into the reservoir (the nominal dose). Hence, less drug wouldneed to be loaded into the pulsating membrane nebulizer to treat thepatient in need thereof to provide the same “dose to subject” asprovided by an air jet nebulizer.

Aerosol droplet size was determined using a Malvern Spraytec instrument.The Dv10, Dv50, and Dv90 from the pulsating membrane nebulizer are verysimilar to those for the reference product (PULMICORT RESPULES) and aCAPTISOL solution of the same concentration in an air jet nebulizer Thissuggests that the formulation would be similarly distributed within thepatient after inhalation.

A clinical study according to Example 17 was conducted to compare thepulmonary disposition of budesonide from a radiolabeled liquidformulation and the clearance of budesonide from the lung as determinedby appearance of budesonide in the plasma. A solution formulation of theinvention was compared to a suspension-based formulation of budesonide.At various times up to 24 hours, plasma samples were collected andassayed for budesonide and various pharmacokinetic parametersdetermined. The area under the plasma concentration—time curve (AUC) isa measure of the delivery of budesonide to the lung, since oralabsorption of the corticosteroid was blocked by the administration ofcharcoal. A comparison of the AUC data was made by consideration of thedose delivered to each subject (“dose to subject”) or dose delivered tothe lungs of each subject (“dose to lung”) or dose emitted by thenebulizer or device (“emitted dose”) or dose available for nebulizationor delivery (“nominal dose” or “nominal available dose” or “loadeddose”). The AUC data (_(0-t), and _(0-∞)) was normalized in terms of thebudesonide (μg) delivered to each subject by dividing the AUC data bythe corresponding dose to subject. As used herein the term “dose tosubject” is taken to mean the amount of corticosteroid delivered to asubject following completion of a dosing cycle with a nebulizer and iscalculated by subtracting the sum of drug remaining in the nebulizer,drug removed from the mouth of a subject, and the amount collected fromthe exhalation filter from the amount of drug initially present in thereservoir of the nebulizer. The following expression can be used tocalculate the dose to subject (Ds): Ds=Dr_(init)−(D_(mw)+D_(dev)),wherein Dr_(init) denotes the amount of drug initially present in thereservoir of the nebulizer, D_(mw) denotes the amount of drug removedfrom the mouth of a subject by using a mouthwash, D_(dev) denotes theamount of drug remaining in the device following completion of anadministration dosing cycle. The term D_(dev) includes drug remaining inthe reservoir after completion of dosing, and drug remaining in theremainder of the device after completion of dosing. As used herein, theterm “dose to lung” is taken to mean the amount of drug delivered to thelungs of a subject, which amount is a subset of “dose to subject”. Thedosing parameters used in this example are detailed below and areprovided on the basis of arithmetic means.

Arithmetic Means Pulmicort Parameter Captisol Enabled BudesonideRespules Normalization or Ratio 25% TTS 50% TTS 75% TTS 100% TTSIntended μg 1000 1000 1000 1000 Nominal Ratio 1 1 1 1 Loaded Dose CEB/PRNominal μg 250 500 750 1000 Available Ratio 0.25 0.50 0.75 1 Dose CEB/PRDose to Lung μg 39.56 63.85 94.38 90.17 Range = Range = Range = Range =24.61→65.65 48.27→80.70 72.81→123.79 76.21→100.75 Ratio 0.44 0.72 1.05 1CEB/PR Range = Range = Range = 0.27→0.78 0.57→0.95 0.87→1.46 Dose to μg64.1 109.1 169.8 199.9 Subject Range = Range = Range = Range =47.57→81.75 85.43→156.67 142.48→219.03 159.58→249.88 Ratio 0.33 0.550.86 1 CEB/PR Range = Range = Range = 0.23→0.48 0.41→0.64 0.74→1.04Emitted Dose μg 119.5 205.9 321.8 440.2 Range = Range = Range = Range =93.40→166.2 164.0→272.9 277.9→391.0 396.7→481.10 Ratio 0.27 0.47 0.73 1CEB/PR Range = Range = Range = 0.20→0.37 0.41→0.57 0.66→0.87

The pharmacokinetic data obtained from the study of Example 17 issummarized below. The data is presented on a geometric mean orindividual subject basis. The data in the table below is based upon the“dose to subject”.

Pulmicort Captisol Enabled Budesonide Respules Geometric Means Parameter25% TTS¹ 50% TTS¹ 75% TTS¹ 100% TTS Dose to Dose to Dose to Dose toSubject = Subject = Subject = Subject = 62.7 μg 107.1 μg 168.2 μg 197.7μg Range³ = Range³ = Range³ = Range³ = 47.6 →81.8 85.4 →156.7 142.5→219.0 159.6 →249.9 Non-normalized AUC_(0-t) 370 875 1130 516 (pg ·h/ml) Range = Range = Range = Range = 162 →1209 647 →1122 865 →1588 378→631 AUC_(0-∞) 533 1022  1617 644 (pg · h/ml) Range = Range = Range=Range = 279 →1309 778 →1285 978 →2351 421 →864 Normalized AUC_(0-t)  6 8   7  3 for dose (pg · h/ml)/μg Range = Range = Range = Range = tosubject 3 →15 7 →11 4 →11 2 →5 AUC_(0-∞)  8  10  10  3 (pg · h/ml)/μgRange = Range = Range = Range = 4 →16 8 →12 5 →16 2 →5 ¹TTS = Time toSputter. The percentage value can be used to calculate a nominalavailable dose for the corticosteroid in the study conducted accordingto Example 17. ²Using the added radioactive tracer, the dose ofbudesonide delivered may be calculated by subtracting the amount ofbudesonide remaining in the nebulizer post-dose, on the exhalationfilter, and in the mouthwash from the amount initially added to thenebulizer. ³The ranges in the table above are based upon the geometricmeans for a respective value as determined with each individual.

Based upon the above data, the formulation of the invention provides anormalized AUCt of 3-15 (or about at least 6) (pg*h/ml)/μg of budesonidewhen about 60-65 μg of budesonide were delivered, a normalized AUCt of7-11 (or about at least 8) (pg*h/ml)/μg of budesonide when about 105-110μg of budesonide were delivered, and a normalized AUCt of 4-11 (or aboutat least 7) (pg*h/ml)/μg of budesonide when 165-170 μg of budesonidewere delivered, based upon the “dose to subject”. In addition, theformulation of the invention provided a normalized AUCi of 4-16 (orabout at least 8) (pg*h/ml)/μg of budesonide when about 60-65 μg ofbudesonide were delivered, a normalized AUCi of 8-12 (or about at least10) (pg*h/ml)/μg of budesonide when about 105-110 μg of budesonide weredelivered, and a normalized AUCi of 5-16 (or about at least 10)(pg*h/ml)/μg of budesonide when 165-170 μg of budesonide were delivered,based upon the “dose to subject”. The data is also summarized in FIG.17, which is a plot of the geometric mean of “dose to subject” (μgbudesonide) versus the geometric mean of AUC (pg*h/ml), and FIG. 18,which is a plot of the geometric mean of “dose to lung” (μg budesonide)versus the geometric mean of AUC (pg*h/ml).

In some embodiments, the invention includes a method of providing in asubject a mean plasma AUCt, normalized for dose of corticosteroid tosubject, of at least 6 (pg*h/ml)/pg of corticosteroid delivered, as doseto subject, comprising: administering to the subject via nebulization aunit dose comprising at least 45 μg, at least 48 μg, or 45 μg to 1000μg, about 1 μg to 20 mg, about 1 μg to 10 mg, 0.01 mg to 10 mg, 0.025 mgto 10 mg, 0.05 mg to 5 mg, 0.1 mg to 5 mg, 0.125 mg to 5 mg, 0.25 mg to5 mg, 0.5 mg to 5 mg, 0.05 mg to 2 mg, 0.1 mg to 2 mg, 0.125 mg to 2 mg,0.25 mg to 2 mg, or 0.5 mg to 2 mg of corticosteroid dissolved in anaqueous liquid carrier comprising sulfoalkyl ether cyclodextrin.

FIG. 19 is a plot of the “dose to subject” (μg budesonide) versus thecorresponding AUC (pg*h/ml) for each individual subject of the study.The slope of the substantially linear solid line, taken from data acrossthree different doses delivered, defines a dose response curve for apatient receiving the corticosteroid. As a result, the slope can be usedto predict the dose a patient would need to provide a target plasmalevel at a second dose if the patient has received a first dose and thepatient's AUC per μg of corticosteroid has been determined. The sloperanges from 5.7 to 16, or more specifically from 9 to 10, when the datais viewed on an individual subject basis

The plasma concentration profile for budesonide for the subjects of theclinical study is depicted in FIG. 20.

This normalized AUC data and associated radiolabel distribution datashow that more of the dose delivered to the subject made it into thelung and from there into systemic circulation when budesonide wasadministered as a solution than when the budesonide was administered asa suspension. Assuming that an equivalent dose deposited in the lungresults in a similar efficacy and systemic absorption, these resultssuggest that 1.6 to 5 times or 2 to 4 times less dose to subject isrequired when administered in solution to be as effective as thereference suspension product. Depending upon whether it is determined onan individual basis or a geometric mean basis, administration of asolution of the invention provides a 1.6-fold increase, 2.2-foldincrease, a 2.5-fold increase, a five-fold increase, a 1.6 to five foldincrease, a two to four-fold increase, a two to 3.5-fold increase, a twoto 3.3-fold increase or at least a two-fold increase in the AUCt or AUCiper μg of budesonide delivered as compared to administration of thePULMICORT RESPULES suspension-based aqueous formulation. The AUCt orAUCi per μg of budesonide delivered was observed as varying perindividual, with the value being higher than the above-noted values forsome individuals and lower than the above-noted values for otherindividuals.

The data in the table below is based upon the various different dosesmeasured or determined during the study and summarizes the level(factor) of enhancement in the pharmacokinetic profile parameter C_(max)non-normalized or normalized on the basis of nominal available dose,dose to lung, dose to subject, or emitted dose.

Geometric Means Pulmicort Parameter Captisol Enabled Budesonide RespulesNormalization or Ratio 25% TTS 50% TTS 75% TTS 100% TTS Non-normalizedC_(max) 225.8   437.2   545.5   266.3  (pg/mL) Range = Range = Range =Range = 91.6→483.3 297.1→739.1 376.6→891.6 155.8→331.5 Ratio 0.85 1.642.05 1 CEB/PR Range = Range = Range = 0.55→1.55 0.90→2.60 1.50→2.86Normalized (pg/mL) 0.90 0.87 0.73   0.27 for nominal μg Range = Range =Range = Range = available dose¹ 0.37→1.93 0.59→1.48 0.50→1.19 0.16→0.33Ratio 3.39 3.28 2.73 1 CEB/PR Range = Range = Range = 2.20→6.211.79→5.20 2.01→3.82 Normalised (pg/mL) 6.03 6.93 5.86   2.97 for dose μgRange = Range = Range = Range = to lung² 3.49→8.51 5.49→9.16 4.52→7.451.59→3.95 Ratio 2.03 2.33 1.97 1 CEB/PR Range = Range = Range =1.48→2.59 1.56→4.30 1.38→3.17 Normalised (pg/mL) 3.60 4.08 3.24   1.35for dose μg Range = Range = Range = Range = to subject³ 1.93→5.943.22→7.44 1.72→5.23 0.75→1.85 Ratio 2.67 3.03 2.41 1 CEB/PR Range =Range = Range = 1.99→3.21 2.17→5.01 1.72→3.76 Normalized (pg/mL) 1.932.15 1.71   0.61 for emitted μg Range = Range = Range = Range = dose⁴0.98→2.91 1.71→3.25 1.00→2.68 0.34→0.84 Ratio 3.18 3.55 2.82 1 CEB/PRRange = Range = Range = 2.19→4.20 2.17→6.26 1.93→4.23

According to the table above, the formulation and system of theinvention can provide a C_(max) (pg/ml) of 90 to 900, 200 to 600, 200 to550, 200 to 250, 400 to 450, 500 to 600, 225, 437, or 545 on a dosenon-normalized basis. The formulation and system of the invention canprovide a dose normalized C_(max) (pg/ml/μg) of: 1) 0.3 to 2, 0.35 to 2,0.6 to 1.5, 0.5 to 1.2, 0.8 to 1, 0.8 to 0.9, 0.7 to 0.8, 0.4, 1.9, 0.6,1.5, 0.5, 1.2, 0.35, 2, 0.7, 0.8, or 0.9 on a nominal available dosenormalized basis; 2) 3.4 to 9.2, 3.5 to 8.5, 5.5 to 9.2, 4.5 to 7.5, 5.8to 7, 3.4, 3.5, 4.5, 5.5, 9.2, 8.5, 7.5, 5.8, 5.9, 6, or 7 on a dose tolung normalized basis; 3) 1.7 to 7.5, 3.2 to 4.1, 1.9 to 6, 3.2 to 7.5,1.7 to 5.2, 3.6, 4.1, 3.2, 1.9, 6, 3.2, 7.4, 7.5, 1.7, 5.2 or 5.3 on adose to subject normalized basis; 4) 0.9 to 3.3, 1.7 to 2.2, 0.9 to 3, 1to 3, 1.7 to 3.3, 1 to 2.7, 1.9, 2.1, 2.2, 1.7, 0.9, 1, 2, 3, 2.9, 3.2,3.3, or 2.7 on an emitted dose normalized basis.

The solution of the invention provides an enhanced pharmacokineticprofile over the suspension-based formulation. On the basis of thenon-normalized dose of corticosteroid, the Cmaxprovided by thecorticosteroid solution is 1.6 to 2, 1.5 to 3, 1.5 to 2.5, 1.5 to 2,1.5, 1, 6, 2, 2.5, or 3 fold higher than the Cmaxprovided by thesuspension-based formulation when the dose of corticosteroid in thesolution and the suspension is approximately the same amount loaded. Onthe basis of normalization to the nominal available dose ofcorticosteroid, the Cmaxprovided by the corticosteroid solution is 1.8to 6.2, 1.5 to 6.5, 2 to 6.5, 1.5 to 5.5, 2 to 4, 1.5 to 4, 1.5 to 3,2.7, 3.3, 3.4, 1.5, 6.5, 2, 5.5, 4, or 3 fold higher than theCmaxprovided by the suspension-based formulation. On the basis ofnormalization to the dose of corticosteroid to lung, the Cmaxprovided bythe corticosteroid solution is 1.4 to 4.3, 1.4 to 4.5, 1.5 to 4.5, 1.5to 3.5, 1.5 to 3, 1.4 to 3, 1.5 to 2.5, 1.5 to 2, 2, 2.3, 1.4, 4.5, 3.5,3, 1.5 or 2.5 fold higher than the Cmaxprovided by the suspension-basedformulation. On the basis of normalization to the dose of corticosteroidto subject, the Cmaxprovided by the corticosteroid solution is 2 to 3.5,2 to 5, 1.7 to 3.8, 1.7 to 5, 2.7, 3, 2.4, 2, 3.5, 5, 1.7, or 3.8 foldhigher than the Cmaxprovided by the suspension-based formulation. On thebasis of normalization to the emitted dose of corticosteroid, theCmaxprovided by the corticosteroid solution is 1.9 to 6.3, 1.75 to 6.5,2.2 to 4.2, 2.2 to 6.3, 1.9 to 4.2, 3.2, 3.5, 3.6, 2.8, 1.75, 6.5, 2.2,4.2, or 6.3 fold higher than the Cmaxprovided by the suspension-basedformulation.

Alternatively, the Cmaxprovided by the corticosteroid solution is atleast 1.5, 1.6, 2, 2.6, and 3 fold higher than the Cmaxprovided by thesuspension-based formulation when the dose of corticosteroid in thesolution is about 2 fold lower than the dose in the suspension.

The data in the table below is based upon the various different dosesmeasured or determined during the study and summarize the level (factor)of enhancement in the pharmacokinetic profile parameter AUC_(inf)non-normalized or normalized on the basis of nominal available dose,dose to lung, dose to subject, or emitted dose.

Geometric Means Pulmicort Parameter Captisol Enabled Budesonide RespulesNormalization or Ratio 25% TTS 50% TTS 75% TTS 100% TTS Non-normalizedAUCinf 533.0   1022.2   1616.8   643.6  (pg*h/mL) Range = Range = Range= Range = 278.6→1309 777.5→1285 977.5→2438.2 420.6→864.1 Ratio 0.83 1.592.51 1 CEB/PR Range = Range = Range = 0.44→1.51 1.00→3.06 1.53→3.28Normalized (pg*h/mL) 2.13 2.04 2.16   0.64 for nominal μg Range = Range= Range = Range = available dose 1.11→5.23 1.55→2.57 1.30→3.25 0.42→0.86Ratio 3.31 3.18 3.35 1 CEB/PR Range = Range = Range = 1.77→6.062.00→6.11 2.04→4.37 Normalized (pg*h/mL) 14.24  16.20  17.37    7.17 fordose μg Range = Range = Range = Range = to lung 10.17→19.94 13.59→18.7711.16→24.69 5.52→10.22 Ratio 1.98 2.26 2.42 1 CEB/PR Range = Range =Range = 1.19→2.85 1.47→3.40 1.37→3.52 Normalized (pg*h/mL) 8.50 9.549.61   3.26 for dose μg Range = Range = Range = Range = to subject4.23→16.08 8.01→12.23 5.41→16.5 1.68→5.14 Ratio 2.61 2.93 2.95 1 CEB/PRRange = Range = Range = 1.60→3.64 2.00→4.87 1.93→3.93 Normalized(pg*h/mL) 4.55 5.04 5.07   1.43 for emitted μg Range = Range = Range =Range = dose 2.62→7.87 4.23→6.66 3.16→8.48 0.81→2.31 Ratio 3.18 3.523.55 1 CEB/PR Range = Range = Range = 1.92→5.37 2.31→5.82 1.91→5.47

According to the table above, the formulation and system of theinvention can provide an AUC_(inf) (pg*h/ml) of 500 to 1700, 530 to1650, 250 to 2500, 280 to 1300, 780 to 1300, 980 to 2450, 275, 775, 980,2400, 2500, 1300, 1290, 530, 1650, 250, 280 or 780 on a dosenon-normalized basis. The formulation and system of the invention canprovide a dose normalized AUC_(inf) (pg/ml/μg) of: 1) 1 to 5.5, 2 to2.2, 1 to 5.3, 1.1 to 5.2, 1.5 to 2.6, 1.3 to 3.3, 2, 2.1, 2.2, 1, 5.5,5.3, 5.2, 1.5, 2.6, 1.3 or 3.3 on a nominal available dose normalizedbasis; 2) 10 to 25, 14 to 18, 10.2 to 20, 13.6 to 18.8, 11.2 to 24.7,10.2, 20, 13.6, 14, 19, 18.8, 11, 11.2, 25, 24.7, 14.2, 16.2, 17.3 on adose to lung normalized basis; 3) 4 to 16, 4.2 to 16.1, 8 to 12.2, 5.4to 16, 5.4 to 17, 8.5 to 9.6, 8.5, 9.5, 9.6, 4.2, 16.1, 8, 12.2, 12,5.4, 16, 17, or 16.5 on a dose to subject normalized basis; 4) 2.5 to 9,2.6 to 8.5, 4.5 to 5.1, 2.5 to 8, 2.6 to 7.9, 4.2 to 6.7, 3.1 to 8.5,3.2 to 8.5, 4.5, 4.6, 5, 5.1, 2.5, 2.6, 4.2, 3.1, 9, 8.5, 5.1, 8, 7.9,or 6.7 to on an emitted dose normalized basis.

Accordingly, the solution of the invention provides an enhancedpharmacokinetic profile over the suspension-based formulation. On thebasis of the non-normalized dose of corticosteroid, the AUC_(inf)provided by the corticosteroid solution is 1.6 to 2.5, 1.6 to 3.1, 1.5to 3.5, 1.5 to 3.3, 2.5 to 3.3, 3.1, 1.5, 3.3, 1.6, or 2.5 fold higherthan the AUC_(inf) provided by the suspension-based formulation when thedose of corticosteroid in the solution and the suspension isapproximately the same. On the basis of normalization to the nominalavailable dose of corticosteroid, the AUC_(inf) provided by thecorticosteroid solution is 1.75 to 6.5, 1.75 to 6.1, 2 to 6.5, 2 to 6.1,2 to 4.5, 2 to 4.4, 3.3, 3.2, 3.5, 3.4, 1.75, 6.5, 6.1, 2, 4.5, or 4.4fold higher than the AUC_(inf) provided by the suspension-basedformulation. On the basis of normalization to the dose of corticosteroidto lung, the AUC_(inf) provided by the corticosteroid solution is 1.2 to3.5, 1.2 to 4, 1.2 to 3, 1.2 to 2.85, 1.5 to 3.5, 1.4 to 3.5, 2, 2.2,2.3, 2.4, 1.2, 3, 4, 2.85, 1.5, 3.5, or 1.4 fold higher than theAUC_(inf) provided by the suspension-based formulation. On the basis ofnormalization to the dose of corticosteroid to subject, the AUC_(inf)provided by the corticosteroid solution is 1.6 to 4.9, 1.5 to 5, 1.6 to5, 1.6 to 3.7, 1.6 to 3.6, 2 to 4.9, 1.9 to 4, 2.6, 1.5, 5, 1.6, 3.7,3.6, 2, 4.9, 1.9, or 4 fold higher than the AUC_(inf) provided by thesuspension-based formulation. On the basis of normalization to theemitted dose of corticosteroid, the AUC_(inf) provided by thecorticosteroid solution is 1.5 to 6, 1.7 to 6, 1.9 to 6, 1.9 to 5.4, 2.3to 5.8, 1.9 to 5.5, 1.9 to 5.8, 1.5, 6, 1.7, 1.9, 5.4, 2.3, 5.8, 5.8,3.2, 3.5, or 3.6 fold higher than the AUC_(inf) provided by thesuspension-based formulation.

Alternatively, the AUC_(inf) provided by the corticosteroid solution isat least 1.5, 1.6, 2, 2.5, 3 and 3.1 fold higher than the AUC_(inf)provided by the suspension-based formulation when the dose ofcorticosteroid in the solution is about 2 fold lower than the dose inthe suspension.

The data in the table below is based upon the various different dosesmeasured or determined during the study and summarize the level (factor)of enhancement in the pharmacokinetic profile parameter AUC_(last)non-normalized or normalized on the basis of nominal available dose,dose to lung, dose to subject, or emitted dose.

Geometric Means Pulmicort Parameter Captisol Enabled Budesonide RespulesNormalization or Ratio 25% TTS 50% TTS 75% TTS 100% TTS Non-normalizedAUCtlast 396.7   874.5   1130     516.2  (pg*h/mL) Range = Range = Range= Range = 162.3→1209 646.7→1122 864.6→1588 377.6→630.9 Ratio 0.77 1.692.19 1 CEB/PR 0.32→1.92 1.14→2.97 1.55→2.93 Normalized (pg*h/mL) 1.591.75 1.51   0.52 for nominal μg 0.65→4.84 1.29→2.24 1.15→2.12 0.38→0.63available dose ¹ Ratio 3.07 3.39 2.92 1 CEB/PR 1.29→7.66 2.28→5.942.07→3.90 Normalized (pg*h/mL) 10.60  13.86  12.14    5.75 for dose μg6.18→18.41 11.12→16.82 9.44→17.96 4.96→7.47 to lung ² Ratio 1.84 2.412.11 1 CEB/PR 1.21→2.49 1.68→3.31 1.53→3.38 Normalized (pg*h/mL) 6.338.16 6.72   2.61 for dose μg 3.34→14.85 6.56→10.74 3.95→11.15 1.51→3.75to subject ³ Ratio 2.42 3.13 2.57 1 CEB/PR 1.40→3.96 2.27→4.74 2.09→3.50Normalized (pg*h/mL) 3.39 4.31 3.54   1.18 for emitted μg 1.74→7.273.46→6.01 2.30→5.72 0.78→1.42 dose ⁴ Ratio 2.88 3.67 3.02 1 CEB/PR1.59→5.19 2.52→5.24 2.36→4.03

A determination of dose distribution in subjects (Example 17) was madeto determine the efficiency of dose deposition to the lungs. The resultsare detailed in the table below and in FIG. 22.

1(A) 2(B) 3(C) 4(D) Lung Scintigraphy (mean values) Dose in lungs 90.1739.56 63.85 94.39 (μg) % wt. in lungs 20.61 32.61 31.30 29.61 % wt. in24.79 21.13 21.68 23.39 oropharyngeal/ stomach region % wt. in 54.6146.26 47.02 47.00 exhalation filter/mouthpiece Mass Balance 100.47100.97 103.21 103.49 1(A) Budesonide Suspension nebulized tosputter—100% TTS (time to sputter) 2(B) Captisol-Enabled ® BudesonideFormulation—25% TTS 3(C) Captisol-Enabled ® Budesonide Formulation—50%TTS 4(D) Captisol-Enabled ® Budesonide Formulation—75% TTS

The data demonstrate that administration of the solution of theinvention resulted in a lower oropharyngeal deposition of thecorticosteroid as compared to administration of the control suspensionformulation. Moreover, the formulation provided at least a 20% to 85%,or 30% to 80%, or at least 30%, at least 40%, at least 56%, at least59%, at least 62% wt. pulmonary deposition of corticosteroid based uponthe emitted dose. The percentage of pulmonary deposition can be furtherincreased by employing a nebulizer capable of providing a higherrespirable fraction and lower MMAD for the nebulized mist. Suitablenebulizers are ultrasonic, vibrating mesh, vibrating cone, vibratingplate nebulizers or those that extrude a liquid formulation through aself-contained nozzle array. For example, the Aradigm AERx pulmonarydelivery systems, the AERx Essence and AERx Ultra, is particularlysuitable for use according to the invention, as it is recognized in theart as providing controlled dose expression, control of generatedaerosol particle size, control of inhaled aerosol particle size, andmanagement of the inhalation and delivery process (Farr et al., DrugDelivery Technology May 2002 Vol. 2, No. 3, 42-44). For example, thePARI eFlow vibrating plate nebulizer is particularly suitable for useaccording to the invention, as it is recognized in the art as providingthe above-mentioned desired performance parameters (Keller et al. (ATS99^(th) International Conference, Seattle, May 16^(th)-21^(st), 2003;poster 2727).

By virtue of the improved systemic bioavailability of a corticosteroiddelivered to the lungs by pulmonary administration according to theinvention, the composition or formulation of the invention will providean improved therapeutic benefit or improved clinical benefit over anequivalent dose of corticosteroid administered as an aqueous suspension.

Example 30 details a study conducted on dogs wherein a budesonidesolution of the invention and a reference formulation (PULMICORTRESPULES suspension) was administered via nebulization with a Pari LCair-jet nebulizer. The results depicted in FIG. 21 demonstrate anaverage plasma level of budesonide as a function of time. TheAUC_(0→8 hr) of the solution formulation was 1.9 times greater than thereference suspension formulation. The solution formulation also emitted1.23 times as much budesonide as the reference formulation in the samelength of time. Detailed results are also provided in the table below.

Dog PK Study Captisol Enabled Pulmicort Ratio Parameter BudesonideRespules CEB/PR Loaded Dose (nominal dose) 1000 μg 1000 μg 1 EmittedDose 769 μg 626 μg 1.23 Average AUC_(0→8 hr) 2599 pg × hr/mL 1355 pg ×hr/mL 1.91 Average C_(max) 1685 pg/mL 901 pg/mL 1.87 LoadedDose—normalized 2.60 (pg × hr/mL)/μg 1.36 (pg × hr/mL)/μg 1.91AUC_(0→8 hr) Emitted Dose—normalized 3.38 (pg × hr/mL)/μg 2.16 (pg ×hr/mL)/μg 1.56 AUC_(0→8 hr) Loaded Dose—normalized 1.69 (pg/mL)/μg 0.90(pg/mL)/μg 1.88 C_(max) Emitted Dose—normalized 2.19 (pg/mL)/μg 1.44(pg/mL)/μg 1.52 C_(max)

According to the table above, the formulation and system of theinvention can provide an AUC_(0-8 hr) (pg*h/ml) of 2000 to 3000, 2500 to2700, 2000, 3000, or 2600 on a dose non-normalized basis. Theformulation and system of the invention can provide a dose normalizedAUC_(inf) (pg/ml/μg) of: 1) 2 to 3, 2.5 to 2.7, or 2.6 on a loaded dosenominal normalized basis; 2) 3 to 4, 3.4 to 3.5, or 3.4 on an emitteddose normalized basis.

The formulation, system and method of the invention provided a higherAUC_(0-8 hr) than did the suspension-based PULMICORT RESPULESformulation. On the basis of the normalized nominal (loaded) dose ofcorticosteroid, the AUC_(0-8 hr) provided by the corticosteroid solutionis at least 1.7, 1.8, 1.9, or 2 fold higher than the AUC_(0-8 hr)provided by the suspension-based formulation when the dose ofcorticosteroid in the solution and the suspension is approximately thesame. On the basis of normalization to the emitted dose ofcorticosteroid, the AUC_(0-8 hr) provided by the corticosteroid solutionis at least 1.5, 1.6, to 2 fold higher than the AUC_(0-8 hr) provided bythe suspension-based formulation.

According to the table above, the formulation and system of theinvention can provide an C_(max) (pg/ml) of 1600 to 1800, 1650 to 1750,or 1700 on a dose non-normalized basis. The formulation and system ofthe invention can provide a dose normalized C_(max) (pg/ml/μg) of: 1) 1to 2, 1.6 to 1.8, or 1.7 on a loaded dose nominal normalized basis; 2)2to 2.5, or 2.2 on an emitted dose normalized basis.

The formulation, system and method of the invention provided a higherC_(max) than did the suspension-based PULMICORT RESPULES formulation. Onthe basis of the normalized nominal (loaded) dose of corticosteroid, theC provided by the corticosteroid solution is at least 1.7, 1.8, 1.9, or2 fold higher than the C_(max) provided by the suspension-basedformulation when the dose of corticosteroid in the solution and thesuspension is approximately the same. On the basis of normalization tothe emitted dose of corticosteroid, the C_(max) provided by thecorticosteroid solution is at least 1.5, 1.6, to 2 fold higher than theC_(max) provided by the suspension-based formulation.

In some aspects, the method and dosage form of the invention thusprovides an improved method of administering a corticosteroidsuspension-based unit dose, the method comprising the step of adding asufficient amount of SAE-CD to convert the suspension to a clearsolution and then administering the clear solution to a subject. As aresult, the method of the invention provides increased total delivery ofthe corticosteroid as well as increased rate of administration ascompared to the initial unit dose suspension formulation.

The data herein can also be used to estimate the percentage of the doseof corticosteroid emitted or delivered that is absorbed into thebloodstream of a patient. In some embodiments, greater than 20%, greaterthan 25%, greater than 29%, greater than 35% of the emitted dose may beabsorbed into the bloodstream of the patient. In some embodiments,greater than 40%, greater than 50%, greater than 55% of the dose tosubject may be absorbed into the bloodstream of the patient. In someembodiments, greater than 10%, greater than 12%, greater than 15% of thenominal available dose may be absorbed into the bloodstream of thepatient.

The corticosteroids that are useful in the present invention generallyinclude any steroid produced by the adrenocortex, includingglucocorticoids and mineralocorticoids, and synthetic analogs andderivatives of naturally occurring corticosteroids havinganti-inflammatory activity. Suitable synthetic analogs include prodrugs,ester derivatives Examples of corticosteroids that can be used in thecompositions of the invention include aldosterone, beclomethasone,betamethasone, budesonide, ciclesonide (Altana Pharma AG), cloprednol,cortisone, cortivazol, deoxycortone, desonide, desoximetasone,dexamethasone, difluorocortolone, fluclorolone, flumethasone,flunisolide, fluocinolone, fluocinonide, fluocortin butyl,fluorocortisone, fluorocortolone, fluorometholone, flurandrenolone,fluticasone, halcinonide, hydrocortisone, icomethasone, meprednisone,methylprednisolone, mometasone, paramethasone, prednisolone, prednisone,rofleponide, RPR 106541, tixocortol, triamcinolone, and their respectivepharmaceutically acceptable derivatives, such as beclomethasonedipropionate (anhydrous or monohydrate), beclomethasone monopropionate,dexamethasone 21-isonicotinate, fluticasone propionate, icomethasoneenbutate, tixocortol 21-pivalate, and triamcinolone acetonide.Particularly preferred are compounds such as beclomethasonedipropionate, budesonide, flunisolide, fluticasone propionate,mometasone furoate, and triamcinolone acetonide. Other corticosteroidsnot yet commercialized, but that are commercialized subsequent to thefiling of this application, are considered useful in the presentinvention unless it is otherwise established experimentally that theyare not suitable.

Corticosteroids can be grouped according to their relative lipophilicityas described by Barnes et al. (Am. J. Respir. Care Med. (1998), 157, p.S1-S53), Miller-Larsson et al. (Am J. Respir. Crit. Care Med. (2003),167, A773), D. E. Mager et al. (J. Pharm. Sci. (November 2002), 91(11),2441-2451) or S. Edsbäcker (Uptake, retention, and biotransformation ofcortico steroids in the lung and airways. In: Schleimer R P, O'Byrne P MO, Szefler S J, Brattsand R, editor(s). Inhaled steroids in asthma:optimizing effects in the airways. New York: Marcel Dekker, 2002:213-246). Generally, the less lipophilic a corticosteroid is, the lowerthe amount of SAE-CD required to dissolve it in an aqueous medium andvice versa. Corticosteroids that are less lipophilic than flunisolidegenerally require a SAE-CD to corticosteroid molar ratio of less than10:1 to dissolve the corticosteroid in an aqueous medium. Exemplarycorticosteroids of this group include hydrocortisone, prednisolone,prednisone, dexamethasone, betamethasone, methylprednisolone,triamcinolone, and fluocortolone. Some embodiments of the inventionexclude corticosteroids that are less lipophilic than flunisolide.

Corticosteroids that are at least as lipophilic as or more lipophilicthan flunisolide generally require a SAE-CD to corticosteroid molarratio of more than 10:1 to dissolve the corticosteroid in an aqueousmedium. In some embodiments, the corticosteroid used in the invention isat least as lipophilic as or more lipophilic than flunisolide. Exemplarycorticosteroids of this group include beclomethasone, beclomethasonedipropionate, beclomethasone monopropionate, budesonide, ciclesonide,desisobutyryl-ciclesonide, flunisolide, fluticasone, fluticasonepropionate, mometasone, mometasone furoate, triamcinolone acetonide.

The suitability of a corticosteroid for use in the inhalable liquidcomposition/formulation can be determined by performing a phasesolubility binding study as detailed in Example 23. Phase solubilitybinding data is used to determine the saturated solubility of acorticosteroid in the presence of varying amounts of SAE-CD in anaqueous liquid carrier. The phase solubility binding curve depicted inFIG. 3 demonstrates the saturated solubility of budesonide in an aqueousliquid carrier comprising γ-CD, HP-β-CD or SBE7-β-CD. A phase solubilitycurve in the graph defines the boundary for the saturated solubility thecorticosteroid in solutions containing various different concentrationsof cyclodextrin. A molar phase solubility curve can be used to determinethe molar ratio of SAE-CD to corticosteroid or of corticosteroid toSAE-CD at various concentrations of corticosteroid. The area below thephase solubility curve, e.g. of FIG. 3, denotes the region where thecorticosteroid is solubilized in an aqueous liquid medium to provide asubstantially clear aqueous solution. In this region, the SAE-CD ispresent in molar excess of the corticosteroid and in an amountsufficient to solubilize the corticosteroid present in the liquidcarrier. The boundary defined by the phase solubility curve will varyaccording to the corticosteroid and SAE-CD within a composition orformulation of the invention. The table below provides a summary of theminimum molar ratio of SAE-CD to corticosteroid required to achieve thesaturated solubility of the corticosteroid in the composition orformulation of the invention under the conditions studied.

Approximate Molar Ratio at Saturated Solubility of Corticosteroid*Corticosteroid SAE-CD (SAE-CD:corticosteroid) Beclomethasone SAE-β-CD358 dipropionate Beclomethasone SAE-γ-CD 62 dipropionate BudesonideSAE-β-CD 16 Budesonide SAE-γ-CD 13 (SBE6.1), 10.8 (SBE5.2), 10.1(SPE5.4) Budesonide SAE-α-CD 12 Flunisolide SAE-β-CD 16 FlunisolideSAE-γ-CD 9 Fluticasone SAE-β-CD 32 Fluticasone SAE-β-CD 797 PropionateFluticasone SAE-γ-CD 51 Propionate Fluticasone SAE-α-CD 501 PropionateMometasone SAE-α-CD 73 Mometasone SAE-β-CD 33 Mometasone SAE-α-CD 141furoate Mometasone SAE-β-CD 274 furoate Mometasone SAE-γ-CD 101 furoateTriamcinolone SAE-β-CD 9 acetonide *This value was determined in thepresence of SAE-CD under the conditions detailed in Examples 18, 23accompanying the solubility values presented in the preceding andfollowing text.

The saturated solubility of a corticosteroid in the presence of a fixedamount of SAE-CD will vary according to the identity of thecorticosteroid and the SAE-CD. The table below summarizes somesolubility data for the listed corticosteroids in the absence (intrinsicsolubility of corticosteroid in the aqueous test medium) and in thepresence of two different SAE-CD's as determined herein.

[Steroid] ×10⁵ M Intrinsic Solubility Captisol (SBE)_(6.1) γ-CD Steroid(In H₂O) (0.04M) (0.04M) Hydrocortisone 92.4 2656.3 2369.3Methylprednisolone 43.6 743.1 1215.3 Prednisolone 62.5 1995.3 2095.0Prednisone 50.5 1832.7 1313.7 Triamcinolone Acetonide 3.56 457.0 1059.5Flunisolide 11.3 261.5 455.1 Budesonide 6.6 254.8 306.6 FluticasonePropionate 0.39 5.41 51.8 Beclomethasone Dipropionate 0.41 11.6 46.8Mometasone Fuorate 1.82 16.4 41.5

The above data can be used in combination with the phase solubility datato prepare formulations according to the invention having a targetconcentration of corticosteroid and SAE-CD. Accordingly, someembodiments of the invention comprise a corticosteroid having anintrinsic solubility in water that approximates or is less than theintrinsic solubility of flunisolide (less than about 11×10⁻⁵ M or lessthan about 11.3×10⁻⁵ M) in water as determined herein.

Even though a composition or formulation of the invention can comprisethe corticosteroid present in an aqueous medium at a concentration up toits saturated solubility in the presence of a particular concentrationof SAE-CD, some embodiments of the invention include those wherein thecorticosteroid is present at a concentration that is less than itssaturated solubility in the presence of that concentration of SAE-CD.The corticosteroid can be present at a concentration that is 95% orless, 90% or less, 85% or less, 80% or less, or 50% or less of itssaturated solubility as determined in the presence of SAE-CD. It isgenerally easier to prepare solutions that comprise the corticosteroidat a concentration that is less than its saturated solubility in thepresence of SAE-CD.

Therefore, the molar ratio of SAE-CD to corticosteroid in a formulationor composition of the invention can exceed the molar ratio obtained atthe saturated solubility of the corticosteroid in the presence ofSAE-CD, such as defined by the phase solubility binding curve for thecorticosteroid. In such a case, the molar ratio of SAE-CD tocorticosteroid in the composition or formulation will be at least 1%, atleast 2%, at least 5%, at least 7.5%, at least 10%, at least 15%, atleast 20%, at least 25%, at least 50%, at least 75%, at least 100%, orat least 200% greater than the molar ratio at the saturated solubilityof the corticosteroid in the presence of SAE-CD. For example, if themolar ratio at the saturated solubility is about 14:1, then the molarratio in the composition or formulation can be at least 14.1:1 (for atleast 1% higher), at least 14.3:1 (for at least 2% higher), at least14.7:1 (for at least 5% higher), at least 15.4:1 (for at least 10%higher), at least 16.1:1 (for at least 15% higher), at least 16.8:1 (forat least 20% higher), at least 17.5:1 (for at least 25% higher), atleast 21:1 (for at least 50% higher), at least 24.5:1 (for at least 75%higher), at least 28:1 (for at least 100% higher), or at least 42:1 (forat least 100% higher).

Changes in the molar ratio of SAE-CD to corticosteroid can have animpact upon the total output of a nebulizer. A study was conducted usinga PARI LC PLUS air jet nebulizer and solutions containing varyingamounts of SAE-CD in 2 ml of 250 μg/ml PULMICORT RESPULES. Eachpreparation was nebulized for until no further sustained vapor wasvisibly being emitted. At the end of each run, the amount of budesonideremaining in the reservoir of the nebulizer was determined. The SAE-CDto corticosteroid molar ratios used were 10:1, 14:1, and 20:1. The dataindicate that increasing the molar ratio resulted in an increase of theamount of budesonide delivered and a decrease in the amount ofbudesonide remaining in the reservoir.

Changes in the molar ratio of SAE-CD to corticosteroid can also have animpact upon the dissolution rate of corticosteroid in an aqueous medium.A study was conducted on a roller mixer having containers with solutionscontaining varying amounts of SAE-CD, e.g. CAPTISOL, and a fixed amountof fluticasone propionate or mometasone furoate. The samples wereprepared by mixing the corticosteroid with a solution containing theSAE-CD in a vortexer for about 30 seconds and then placing thecontainers on the roller mixer. Aliquots were taken periodically fromeach container and the amount of dissolved corticosteroid wasdetermined. The SAE-CD to corticosteroid molar ratios used were 10:1,14:1, and 20:1. The data indicate that increasing the molar ratioresulted in an increase in the rate of dissolution of thecorticosteroid.

The corticosteroid compound is present in the final, dilutedcorticosteroid composition designed for inhalation in an amount fromabout 1 μg/ml to about 10 mg/ml, about 10 μg/ml to about 1 mg/ml, orabout 20 μg/ml to about 500 μg/ml. For example, the drug concentrationcan be between about 30 and 1000 μg/ml for triamcinolone acetonide, andbetween about 50 and 2000 μg/ml for budesonide, depending on the volumeto be administered. By following the preferred methods of the presentinvention, relatively high concentrations of the corticosteroid can beachieved in an aqueous-based composition.

Similarly, the corticosteroid compound is present in the final, dilutedcorticosteroid composition designed for nasal administration in anamount from about 10 μg/ml to 6 mg/ml, 50 μg/ml to about 10 mg/ml, about100 μg/ml to about 2 mg/ml, or about 300 μg/ml to about 1 mg/ml. Forexample, the drug concentration can be between about 250 μg/ml and 1mg/ml or 250 μg/ml and 6 mg/ml for triamcinolone acetonide, and betweenabout 400 μg/ml and 1.6 mg/ml or 250 μg/ml and 6 mg/ml for budesonide,depending on the volume to be administered.

For the treatment of pulmonary disorders and sinus-related disorders,the diluted corticosteroid composition is prepared as described herein.The corticosteroid for such treatment is preferably, beclomethasonedipropionate, beclomethasone monopropionate, betamethasone, budesonide,ciclesonide, desisobutyryl-ciclesonide, flunisolide, fluticasone,fluticasone propionate, fluticasone furoate, mometasone, mometasonefuroate, or triamcinolone acetonide, and is formulated in theconcentrations set forth herein. The daily dose of the corticosteroid isgenerally about 0.05 to 10 mg, depending on the drug and the disease, inaccordance with the Physician's Desk Reference (PDR). However, in viewof the improved bioavailability of a corticosteroid when administered asa solution of the invention, the dose required to achieve a desiredclinical endpoint, clinical benefit or therapeutic benefit may be lowerthan the corresponding dose indicated in the PDR.

A unit dose of budesonide may also be administered once daily, onceevery two days, once every week, once every month, or even lessfrequently as set forth in U.S. Pat. No. 6,598,603 and U.S. Pat. No.6,899,099, wherein a dose comprises 0.05 to 2.0 mg or 0.25 to 1.0 mg ofbudesonide. Administration can be during the daytime and/or nighttime. Adose of budesonide, or corticosteroid, can be administered twice, thriceor more times per day or on an as-needed basis.

in some embodiments, a dose comprises 1 μg to 20 mg, 0.01 mg to 10 mg,0.025 mg to 10 mg, 0.05 mg to 5 mg, 0.1 mg to 5 mg, 0.125 mg to 5 mg,0.25 mg to 5 mg, 0.5 mg to 5 mg, 0.05 mg to 2 mg, 0.1 mg to 2 mg, 0.125mg to 2 mg, 0.25 mg to 2 mg, 0.5 mg to 2 mg, 1 μg, 10 μg, 25 μg, 50 μg,100 μg, 125 μg, 200 μg, 250 μg, 25 to 66 μg, 48 to 81 μg, 73 to 125 μg,40 μg, 64 μg, 95 μg, 35 to 95 μg, 25 to 125 μg, 60 to 170 μg, 110 μg,170 μg, 45 to 220 μg, 45 to 85 μg, 48 to 82 μg, 85 to 160 μg, 140 to 220μg, 120 to 325 μg, 205 μg, 320 μg, 325 μg, 90 to 400 μg, 95 to 170 μg,165 to 275 μg, or 275 to 400 μg of budesonide, said dose being a nominaldose, nominal available dose, emitted dose, delivered dose, dose tosubject, or dose to lung.

The corticosteroid can be present in its neutral, ionic, salt, basic,acidic, natural, synthetic, diastereomeric, isomeric, isomeric,enantiomerically pure, racemic, solvate, anhydrous, hydrate, chelate,derivative, analog, esterified, non-esterfied, or other common form.Whenever an active agent is named herein, all such forms available areincluded. For example, all known forms of budesonide are consideredwithin the scope of the invention.

The formulation of the invention can be used to deliver two or moredifferent active agents (active ingredients, therapeutic agents, etc.).Particular combinations of active agents can be provided by the presentformulation. Some combinations of active agents include: 1) a first drugfrom a first therapeutic class and a different second drug from the sametherapeutic class; 2) a first drug from a first therapeutic class and adifferent second drug from a different therapeutic class; 3) a firstdrug having a first type of biological activity and a different seconddrug having about the same biological activity; 4) a first drug having afirst type of biological activity and a different second drug having adifferent second type of biological activity. Exemplary combinations ofactive agents are described herein.

A corticosteroid, such as budesonide, can be administered in combinationwith one or more other drugs (active ingredients, therapeutic agents,active agents, etc., the terms being used interchangeably herein unlessotherwise specified). Such other drugs include: B₂ adrenoreceptoragonist, topical anesthetic, D₂ receptor agonist, anticholinergic agent.

B₂-Adrenoreceptor agonists for use in combination with the compositionsprovided herein include, but are not limited to, Albuterol(alpha¹-(((1,1-dimethylethyl)amino)methyl)-4-hydroxy-1,3-benzenedimethanol);Bambuterol (dimethylcarbamic acid5-(2-((1,1-dimethylethyl)amino)-1-hydroxyethyl)-1,3-phenylene ester);Bitolterol (4-methylbenzoic acid4-(2-((1,1-dimethylethyeamino)-1-hydroxyethyl)-1,2-phenyleneester);Broxaterol(3-bromo-alpha-(((1,1-dimethylethyl)amino)methyl)-5-isoxazolemethanol);Isoproterenol(4-(1-hydroxy-2-((1-methylethyl-)amino)ethyl)-1,2-benzene-diol);Trimetoquinol(1,2,3,4-tetrahydro-1-((3,4-,5-trimethoxyphenyl)-methyl)-6,7-isoquinolinediol);Clenbuterol(4-amino-3,5-dichloro-alpha-(((1,1-diemthylethyl)amino)methyl)benzenemethanol);Fenoterol(5-(1-hydroxy-2-((2-(4-hydroxyphenyl)-1-methylethyl)amino)ethyl)-1,3-benzenediol);Formoterol(2-hydroxy-5-((1RS)-1-hydroxy-2-(((1RS)-2-(p-methoxyphenyl)-1-methylethyl)amino)ethyl)formanilide); (R,R)-Formoterol; Desformoterol ((R,R) or(S,S)-3-amino-4-hydroxy-alpha-(((2-(4-methoxyphenyl)-1-methyl-ethyl)amino)methyl)benzenemethanol);Hexoprenaline(4,4′-(1,6-hexane-diyl)-bis(imino(1-hydroxy-2,1-ethanediyl)))bis-1,2-benzenediol);Isoetharine(4-(1-hydroxy-2((1-methylethyl)-amino)butyl)-1,2-benzenediol);Isoprenaline(4-(1-hydroxy-2-((1-methylethyl)amino)ethyl)-1,2-benzenediol);Meta-proterenol(5-(1-hydroxy-2((1-methylethyl)amino)ethyl)-1,3-benzened-iol);Picumeterol(4-amino-3,5-dichloro-alpha-(((6-(2-(2-pyridinyl)ethoxy)hexyl)-amino)methyl)benzenemethanol);Pirbuterol(.alpha.⁶-(((1,1-dimethylethyl)-amino)methyl)-3-hydroxy-2,6-pyridinemethanol);Procaterol(((R*,S*)-(.+−.)-8-hydroxy-5-(1-hydroxy-2-((1-methylethyl)amino)butyl)-2(1H)-quinolin-one);Reproterol((7-(3-((2-(3,5-dihydroxyphenyl)-2-hydroxyethyl)amino)-propyl)-3,7-dihydro-1,3-dimethyl-1H-purine-2,6-dione);Rimiterol (4-(hydroxy-2-piperidinylmethyl)-1,2-benzenediol); Salbutamol((.+−.)-alpha¹-(((1,1-dimethylethyl)amino)methyl)-4-hydroxy-1,3-b-enzenedimethanol);(R)-Salbutamol; Salmeterol((.+−.)-4-hydroxy-.alpha¹-(((6-(4-phenylbutoxy)hexyl)-amino)methyl)-1,3-benzenedimethanol);(R)-Salmeterol; Terbutaline(5424(1,1-dimethylethyl)amino)-1-hydroxyethyl)-1,3-benzenediol);Tulobuterol(2-chloro-.alpha.-(((1,1-dimethylethyl)amino)methyl)benzenemethanol);and TA-2005(8-hydroxy-5((1R)-1-hydroxy-2-(N-((1R)-2-(4-methoxyphenyl)-1-methylethyl)amino)ethyl)carbostyrilhydrochloride).

Dopamine (D₂) receptor agonists include, but are not limited to,Apomorphine((r)-5,6,6a,7-tetrahydro-6-methyl-4H-dibenzo[de,glquinoline-10,11-diol);Bromocriptine((5′,alpha.)-2-bromo-12′-hydroxy-2′-(1-methylethyl)-5′-(2-methylpropyl)ergotaman-3′,6″,18-trione); Cabergoline((8.beta.)-N-(3-(dimethylamino)propyl)-N-((ethylamino)carbony-1)-6-(2-propenyl)ergoline-8-carboxamide);Lisuride(N′-((8-alpha-)-9,10-didehydro-6-methylergolin-8-yl)-N,N-diethylurea);Pergolide ((8-beta-)-8-((methylthio)methyl)-6-propylergoline); Levodopa(3-hydroxy-L-tryrosine); Pramipexole((s)-4,5,6,7-tetrahydro-N⁶-prop-yl-2,6-benzothiazolediamine); Quinpirolehydrochloride(trans-(−)-4aR-4,4a,5,6,7,8,8a,9-octahydro-5-propyl-1H-pyrazolo[3,4-g]quinolinehydrochloride); Ropinirole(4-(2,-(dipropylamino)ethyl)-1,3-dihydro-2H-indol-2-one); and Talipexole(5,6,7,8-tetrahydro-6-(2-propenyl)-4H-thia-zolo[4,5-d]azepin-2-amine).Other dopamine D₂ receptor agonists for use herein are disclosed inInternational Patent Application Publication No, WO 99/36095, therelevant disclosure of which is hereby incorporated by reference.

Anticholinergic agents for use herein include, but are not limited to,ipratropium bromide, oxitropium bromide, atropine methyl nitrate,atropine sulfate, ipratropium, belladonna extract, scopolamine,scopolamine methobromide, homatropine methobromide, hyoscyamine,isopriopramide, orphenadrine, benzalkonium chloride, tiotropium bromideand glycopyrronium bromide, in certain embodiments, the compositionscontain an anticholinergic agent, such as ipratropium bromide ortiotropium bromide, at a concentration of about 5 μg/mL to about 5mg/mL, or about 50 μg/mL to about 200 μg/mL. In other embodiments, thecompositions for use in the methods herein contain an anticholinergicagent, including ipratropium bromide and tiotropium bromide, at aconcentration of about 83 μg/mL or about 167 μg/mL.

Other active ingredients for use herein in combination therapy, include,but are not limited to, IL-5 inhibitors such as those disclosed in U.S.Pat. No. 5,668,110, U.S. Pat. No. 5,683,983, U.S. Pat. No. 5,677,280,U.S. Pat. No. 6,071,910 and U.S. Pat. No. 5,654,276, the relevantdisclosures of which are hereby incorporated by reference; antisensemodulators of IL-5 such as those disclosed in U.S. Pat. No. 6,136,603,the relevant disclosure of which is hereby incorporated by reference;milrinone (1,6-dihydro-2-methyl-6-oxo-[3,4′-bipyridine]-5-carbonitrile);milrinone lactate; tryptase inhibitors such as those disclosed in U.S.Pat. No. 5,525,623, the relevant disclosure of which is herebyincorporated by reference; tachykinin receptor antagonists such as thosedisclosed in U.S. Pat. No. 5,691,336, U.S. Pat. No. 5,877,191, U.S. Pat.No. 5,929,094, U.S. Pat. No. 5,750,549 and U.S. Pat. No. 5,780,467, therelevant disclosures of which are hereby incorporated by reference;leukotriene receptor antagonists such as montelukast sodium (Singular™.,R-(E)]-1-[[[1-[3-[2-(7-chloro-2-quinolinyl)ethenyl-]phenyl]-3-[2-(1-hydroxy-1-methylethyl)phenyl]-propyl]thio]methyl]cyclopro-paneaceticacid, monosodium salt), 5-lypoxygenase inhibitors such as zileuton(Zyflo™, Abbott Laboratories, Abbott Park, Ill.), and anti-IgEantibodies such as Xolair™ (recombinant humanized anti-IgE monoclonalantibody (CGP 51901; IGE 025A; rhuMAb-E25), Genentech, Inc., South SanFrancisco, Calif.), and topical anesthetics such as lidocaine,N-arylamide, aminoalkylbenzoate, prilocalne, etidocaine (U.S. Pat. No.5,510,339, U.S. Pat. No. 5,631,267, and U.S. Pat. No. 5,837,713, therelevant disclosures of which are hereby incorporated by reference).

Exemplary combination formulations of the invention comprise thefollowing components.

FORM. Corticosteroid (A) Other Active Ingredient (B) I BudesonideFormoterol II Budesonide Salmeterol III Budesonide Albuterol IVFluticasone Salmeterol propionate

A formulation comprising a corticosteroid and another active ingredientcan be prepared according to the examples below. In one embodiment, theSAE-CD is present in an amount sufficient to solubilize thecorticosteroid and the other active ingredient. In another embodiment,the SAE-CD is present in an amount sufficient to solubilize thecorticosteroid or the other active ingredient.

Depending upon the other active ingredient used, it may or may not bindcompetitively against the corticosteroid with the SAE-CD. In someembodiments, the SAE-CD has a higher equilibrium binding constant forthe other active ingredient than it has for the corticosteroid. In someembodiments, the SAE-CD has a higher equilibrium binding constant forthe corticosteroid than it has for the other active ingredient. In someembodiments, the SAE-CD has approximately the same equilibrium bindingconstant for the other active ingredient as it has for thecorticosteroid. Alternatively, the other active ingredient does not bindwith the SAE-CD even though the corticosteroid does. Accordingly, theinvention provides embodiments wherein, the SAE-CD solubilizes thecorticosteroid, the other active ingredient, or a combination thereof.The invention also provides embodiments wherein, the SAE-CD solubilizesat least a major portion of the corticosteroid, the other activeingredient, or of each. The invention also provides embodiments wherein,the SAE-CD does not solubilize the other active ingredient.

The molar ratio of SAE-CD to corticosteroid and SAE-CD to other activeingredient can vary as needed to provide a combination formulation asdescribed herein. The SAE-CD is generally present in molar excess overthe corticosteroid, the other active ingredient, or both.

The phase solubility binding curve for Salmeterol xinafoate andbudesonide was determined as described herein, and the approximateequilibrium binding constant (Ki) of each with CAPTISOL was determined.The approximate Ki for Salmeterol xinafoate was approximately 3,500, andthe approximate Ki for budesonide was 600 under the test conditionsused. The molar ratio of CAPTISOL to Salmeterol xinafoate at saturatedsolubility was about 3.2 under the test conditions used. Their may be areduction in the amount of budesonide solubilized by CAPTISOL in thepresence of Salmeterol xinafoate; however, clear aqueous liquid solutionformulations comprising therapeutically effective amounts of budesonideand Salmeterol xinafoate were prepared.

A formulation comprising budesonide, albuterol and SAE-CD was preparedaccording to Example 25. The formulation was clear after preparation.The combination of budesonide and albuterol has been shown to bephysically and chemically stable and would be expected to provide thesame improved aerosol performance and AUC per μg budesonide dosed as thesolution of budesonide alone. Furthermore, the patient would benefitfrom the simultaneous administration of the two drugs.

The invention includes methods for the treatment, prevention, oramelioration of one or more symptoms of a corticosteroid-responsivedisorder, a disease or disorder of the air passageways e.g. respiratoryor pulmonary disorders such as bronchoconstrictive disorders; sinusdisorders such as sinusitis. The method further includes administeringone or more of (a), (b), (c) or (d) as follows: (a) a b₂-adrenoreceptoragonist; (b) a dopamine (D₂) receptor agonist; (c) a prophylactictherapeutic, such as a steroid; or (d) an anticholinergic agent;simultaneously with, prior to or subsequent to the composition providedherein.

Embodiments of the present invention allow for combinations to beprepared in a variety of ways:

1) Mixing ready to use solutions of a β2-agonist such as levalbuterol oranticholinergic such as ipatropium bromide with a ready to use solutionof a corticosteroid in SAE-CD;

2) Mixing ready to use solutions of a β2-agonist or anticholinergic witha concentrated solution of a corticosteroid dissolved using SAE-CD;

3) Mixing a ready to use solution of a β2-agonist or anticholinergicwith substantially dry SAE-CD and a substantially dry corticosteroid;

4) Mixing a ready to use solution of a β2-agonist or anticholinergicwith a substantially dry mixture of SAE-CD and a corticosteroid or moreconveniently a pre-measured amount of the mixture in a unit containersuch as a capsule (empty a capsule into ready to use solution);

5) Mixing a ready to use solution of a corticosteroid such as budesonidewith a substantially dry long acting or short acting β2-agonist and/orwith a substantially dry anticholinergic such as ipatropium bromide ortiotropium bromide; or

6) Dissolving a substantially dry β2-agonist, and/or a substantially dryanticholinergic and a substantially dry SAE-CD plus a substantially drycorticosteroid.

The materials used herein can be used in micronized or non-micronizedform and crystalline, polymorphic or amorphous form. This isparticularly true of the corticosteroids and other active ingredients.

It is well understood by those of ordinary skill in the art that theabove solutions or powders may optionally contain other ingredients suchas buffers and/or tonicity adjusters and/or antimicrobials and/oradditives or other such excipients as set forth herein or as presentlyused in inhalable liquid formulations to improve the output of thenebulizer.

Dosing, use and administration of the therapeutic agents disclosedherein is generally intended to follow the guidelines set forth in thePhysician's Desk Reference, 55^(th) Edition (Thompson Healthcare,Montvale, N.J., 2005) the relevant disclosure of which is herebyincorporated by reference.

The bronchoconstrictive disorder to be treated, prevented, or whose oneor more symptoms are to be ameliorated is associated with asthma,including, but not limited to, bronchial asthma, allergic asthma andintrinsic asthma, e.g., late asthma and airway hyper-responsiveness;and, particularly in embodiments where an anticholinergic agent is used,other chronic obstructive pulmonary diseases (COPDs), including, but notlimited to, chronic bronchitis, emphysema, and associated cor pulmonale(heart disease secondary to disease of the lungs and respiratory system)with pulmonary hypertension, right ventricular hypertrophy and rightheart failure. COPD is frequently associated with cigarette smoking,infections, environmental pollution and occupational dust exposure.

A formulation according to the invention will generally have a storageshelf life of no less than 6 months. In this case, shelf life isdetermined only as regards the increase in the amount of corticosteroiddegradation by-products or a reduction in the amount of corticosteroidremaining in the formulation. For example, for a formulation having ashelf life of at least six months, the formulation will not demonstratean unacceptable and substantial increase in the amount of degradantsduring the storage period of at least six months. The criteria foracceptable shelf-life are set as needed according to a given product andits storage stability requirements. In other words, the amount ofdegradants in a formulation having an acceptable shelf-life will notincrease beyond a predetermined value during the intended period ofstorage. On the other hand, the amount of degradants of a formulationhaving an unacceptable shelf-life will increase beyond the predeterminedvalue during the intended period of storage.

The method of Example 3 was followed to determine the stability ofbudesonide in solution. The shelf-life was defined as the time to lossof 10% potency. Under the conditions tested, the loss of potency wasfirst order. The shelf life of a Captisol-Enabled® Budesonide InhalationSolution (a solution comprising budesonide and SBE7-β-CD) is greaterthan about 3 years at a pH between 4 and 5, i.e. about 90 months at pH4.0 and about 108 months at pH 5.0 without the need to add any otherstabilizers, such as EDTA, in water in the presence of about 5% wt./vol.SAE-CD. This shelf-life is greater than that reported by Otterbeck (U.S.Pat. No. 5,914,122; up to six weeks at pH 4.0-6.0 in water in thepresence of EDTA, HP-β-CD and other additives.)

The inventors have also discovered that SAE-CD is capable of stabilizingthe isomers of budesonide to different extents. A study to determine ifSBE7-β-CD stabilized budesonide solutions and if it preferentiallystabilized one isomer was conducted according to Example 13. FIG. 11 isa semi-log plot of the % of initial concentration at each time point forthe samples stored at 60° C. Loss of budesonide was first order at eachtemperature. The table below shows the pseudo-first order rate constantscalculated for each isomer at 60° C. and 80° C.

Pseudo 1^(st) Order Rate constant (hours⁻¹) Temperature 60° C. R/SWith/without With/without rate Rate CAPTISOL Rate CAPTISOL con- constantratio for constant ratio for stant Ph R-isomer R-isomers S-isomerS-isomers ratio 4 w/ 0.000597 0.547 0.00012 0.323 5.06 CAPTISOL 4 no0.00109 0.0037 2.99 CAPTISOL 6 w/ 0.001661 0.385 0.000361 0.193 4.60CAPTISOL 6 no 0.00432 0.001872 2.31 CAPTISOL

Pseudo 1^(st) Order Rate constant (hours⁻¹) Temperature 80° C. R/SWith/without With/without rate Rate CAPTISOL Rate CAPTISOL con- Constantratio for constant ratio for stant pH R-isomer R-isomers S-isomerS-isomers ratio 4 w/ 0.002250 0.607 0.000644 0.491 3.49 CAPTISOL 4 no0.003704 0.00131 2.83 CAPTISOL 6 w/ 0.00732 0.529 0.00254 0.384 2.88CAPTISOL 6 no 0.0138 0.00661 2.09 CAPTISOL

SBE7-β-CD stabilized both R- and S-isomers of budesonide in solutions atboth pH 4 and 6. The with/without CAPTISOL ratio of rate constants wasmuch less than 1 at all temperatures. SBE7-β-CD had a greater effect onthe stability of both the R and S-isomer at pH 6 than at pH 4. At agiven temperature the ratio of rate constants with/without SBE7-β-CD wasless at pH 6 than at pH 4. Although SBE7-β-CD stabilized both isomers,the S-isomer appears to be stabilized to an even greater extent than theR. At all temperatures and pHs tested, the ratio of rate constantswith/without SBE7-13-CD was lower for the S isomer. The degree ofstabilization affected by SBE7-P-CD at 60° C. is greater than at 80° C.An even greater degree of stabilization would be expected at 40° C.and/or room temperature (20-30 C).

Samples of the above solutions were also placed in a chamber under abank of fluorescent lights. Vials were periodically removed and assayedfor budesonide. FIG. 12 shows the semi-log plot of the % of initialvalue remaining as a function of light exposure (light intensity*time).As noted in the table below, SBE7-β-CD significantly reduced thephotodecomposition of budesonide. The loss of budesonide was first orderand independent of pH.

Light Stability of Budesonide Pseudo 1st Order Rate constant (hour⁻¹) pH4 pH 6 Captisol 0.0585 0.0562 No Captisol 0.0812 0.0822

The formulation of the invention can be provided as a kit adapted toform an inhalable solution for nebulization. The kit can comprise acorticosteroid, SAE-CD, an aqueous carrier, and optionally one or moreother components. The corticosteroid and SAE-CD can be provided togetheror separately in solid, suspended or dissolved form. After mixing SAE-CDwith corticosteroid in the presence of an aqueous carrier, the solidswill dissolve to form an inhalable solution rather than suspension fornebulization. Each component can be provided in an individual containeror together with another component. For example, SAE-CD can be providedin an aqueous solution while budesonide is provided in dry solid form orwet suspended form. Alternatively, SAE-CD is provided in dry form andbudesonide is provided as an aqueous suspension, e.g., PULMICORTRESPULES™. The kit can instead comprise an admixture of a solidderivatized cyclodextrin and solid cortico steroid and, optionally, atleast one solid pharmaceutical excipient, such that a major portion ofthe active agent is not complexed with the derivatized cyclodextrinprior to reconstitution of the admixture with an aqueous carrier.Alternatively, the composition can comprise a solid mixture comprisingthe inclusion complex of a derivatized cyclodextrin and an active agent,wherein a major portion of the active agent is complexed with thederivatized cyclodextrin prior to reconstitution of the solid mixturewith an aqueous carrier. Depending upon the storage temperature of thekit, the aqueous carrier may be a liquid or frozen solid. In oneembodiment, the kit excludes the aqueous carrier during storage, but theaqueous carrier is added to the SAE-CD and corticosteroid prior to useto form the nebulization solution. The corticosteroid and SAE-CD can becomplexed and present in aqueous concentrated form prior to addition ofthe aqueous carrier, which is later added to bring the solution tovolume and proper viscosity and concentration for nebulization. Areconstitutable formulation can be prepared according to any of thefollowing processes. A liquid formulation of the invention is firstprepared, then a solid is formed by lyophilization (freeze-drying),spray-drying, spray freeze-drying, antisolvent precipitation, variousprocesses utilizing supercritical or near supercritical fluids, or othermethods known to those of ordinary skill in the art to make a solid forreconstitution. Example 29 details a method for the preparation of alyophilized solid composition comprising corticosteroid and SAE-CD bylyophilization of a liquid composition or formulation of the invention.The lyophilized solid can be dissolved in an aqueous liquid carrierprior to administration via nebulization. The dried powder would providea stable form for long-term storage and would also be useful to rapidlyprepare inhalation compositions on a larger scale, or as an additive toanother inhalation solution medication to prepare combination products.

While the liquid composition or formulation of the invention can beadministered to the lung, it would also be suitable for nasal, oral,ophthalmic, otic or topical administration. The liquid composition orformulation may also be administered via inhalation using a device suchas a pump spray, metered dose inhaler, or pressurized metered doseinhaler. Accordingly, the invention provides a method of treating acorticosteroid-responsive disease or disorder by administration of theliquid to a subject in need of such treatment.

A liquid vehicle (carrier) included in a formulation of the inventioncomprises an aqueous liquid carrier, such as water, aqueous alcohol,propylene glycol, or aqueous organic solvent. Example 30 details thepreparation of a liquid formulation comprising 20% w/v SAE-CD,corticosteroid, water and ethanol (0-5%). Increasing the concentrationof the ethanol in the liquid resulted in a decrease in the maximumsaturated solubility of the corticosteroid.

Although not necessary, the formulation of the present invention mayinclude a conventional preservative, antioxidant, buffering agent,acidifying agent, alkalizing agent, colorant, solubility-enhancingagent, complexation-enhancing agent, electrolyte, glucose, stabilizer,tonicity modifier, bulking agent, antifoaming agent, oil, emulsifyingagent, cryoprotectant, plasticizer, flavors, sweeteners, a tonicitymodifier, surface tension modifier, viscosity modifier, densitymodifier, volatility modifier, other excipients known by those ofordinary skill in the art for use in preserved formulations, or acombination thereof.

As used herein, the term “alkalizing agent” is intended to mean acompound used to provide alkaline medium, such as for product stability.Such compounds include, by way of example and without limitation,ammonia solution, ammonium carbonate, diethanolamine, monoethanolamine,potassium hydroxide, sodium borate, sodium carbonate, sodiumbicarbonate, sodium hydroxide, triethanolamine, diethanolamine, organicamine base, alkaline amino acids and trolamine and others known to thoseof ordinary skill in the art.

As used herein, the term “acidifying agent” is intended to mean acompound used to provide an acidic medium for product stability. Suchcompounds include, by way of example and without limitation, aceticacid, acidic amino acids, citric acid, fumaric acid and other alphahydroxy acids, hydrochloric acid, ascorbic acid, phosphoric acid,sulfuric acid, tartaric acid and nitric acid and others known to thoseof ordinary skill in the art.

Inclusion of a conventional preservative in the inhalable solutionformulation is optional, since the formulation is self-preserved bySAE-CD depending upon its concentration in solution. Nonetheless, aconventional preservative can be further included in the formulation ifdesired. Preservatives can be used to inhibit microbial growth in thecompositions. The amount of preservative is generally that which isnecessary to prevent microbial growth in the composition for a storageperiod of at least six months. As used herein, a conventionalpreservative is a compound used to at least reduce the rate at whichbioburden increases, but preferably maintains bioburden steady orreduces bioburden after contamination has occurred. Such compoundsinclude, by way of example and without limitation, benzalkoniumchloride, benzethonium chloride, benzoic acid, benzyl alcohol,cetylpyridinium chloride, chlorobutanol, phenol, phenylethyl alcohol,phenylmercuric nitrate, phenylmercuric acetate, thimerosal, metacresol,myristylgamma picolinium chloride, potassium benzoate, potassiumsorbate, sodium benzoate, sodium propionate, sorbic acid, thymol, andmethyl, ethyl, propyl or butyl parabens and others known to those ofordinary skill in the art.

As used herein, the term “antioxidant” is intended to mean an agent thatinhibits oxidation and thus is used to prevent the deterioration ofpreparations by the oxidative process. Such compounds include, by way ofexample and without limitation, acetone, potassium metabisulfite,potassium sulfite, ascorbic acid, ascorbyl palmitate, citric acid,butylated hydroxyanisole, butylated hydroxytoluene, hypophosphorousacid, monothioglycerol, propyl gallate, sodium ascorbate, sodiumcitrate, sodium sulfide, sodium sulfite, sodium bisulfite, sodiumformaldehyde sulfoxylate, thioglycolic acid, EDTA, pentetate, and sodiummetabisulfite and others known to those of ordinary skill in the art.

As used herein, the term “buffering agent” is intended to mean acompound used to resist change in pH upon dilution or addition of acidor alkali. Buffers are used in the present compositions to adjust the pHto a range of between about 2 and about 8, about 3 to about 7, or about4 to about 5. Such compounds include, by way of example and withoutlimitation, acetic acid, sodium acetate, adipic acid, benzoic acid,sodium benzoate, boric acid, sodium borate, citric acid, glycine, maleicacid, monobasic sodium phosphate, dibasic sodium phosphate, HEPES,lactic acid, tartaric acid, potassium metaphosphate, potassiumphosphate, monobasic sodium acetate, sodium bicarbonate, tris, sodiumtartrate and sodium citrate anhydrous and dihydrate and others known tothose of ordinary skill in the art. Other buffers include citricacid/phosphate mixture, acetate, barbital, borate, Britton-Robinson,cacodylate, citrate, collidine, formate, maleate, Mcllvaine, phosphate,Prideaux-Ward, succinate, citrate-phosphate-borate (Teorell-Stanhagen),veronal acetate, MES (2-(N-morpholino)ethanesulfonic acid), BIS-TRIS(bis(2-hydroxyethyl)imino-tris(hydroxymethyl)methane), ADA(N-(2-acetamido)-2-iminodiacetic acid), ACES(N-(carbamoylmethyl)-2-aminoethanesulfonaic acid), PIPES(piperazine-N,N′-bis(2-ethanesulfonic acid)), MOPSO(3-(N-morpholino)-2-hydroxypropanesulfonic acid), BIS-TRIS PROPANE(1,3-bis(tris(hydroxymethyl)methylamino)propane), BES(N,N-bis(2-hydroxyethyl)-2-aminoethanesulfonaic acid), MOPS(3-(N-morpholino)propanesulfonic acid), TES(N-tris(hydroxymethyl)methyl-2-aminoethanesulfonic acid), HEPES(N-(2-hydroxyethyl)piperazine-N′-(2-ethanesulfonic acid), DIPSO(3-(N,N-bis(2-hydroxyethyl)amino)-2-hydroxypropanesulfonic acid), MOBS(4-(N-morpholino)-butanesulfonic acid), TAPSO(3-(N-tris(hydroxymethyl)methylamino)-2-hydroxypropanesulfonic acid),TRIZMA™ (tris(hydroxymethylaminomethane), HEPPSO(N-(2-hydroxyethyl)piperazine-N′-(2-hydroxypropanesulfonic acid), POPSO(piperazine-N,N′-bis(2-hydroxypropanesulfonic acid)), TEA(triethanolamine),EPPS(N-(2-hydroxyethyl)piperazine-N′-(3-propanesulfonic acid), TRICINE(N-tris(hydroxymethyl)methylglycine), GLY-GLY (glycylglycine), BICINE(N,N-bis(2-hydroxyethyl)glycine), HEPBS(N-(2-hydroxyethyl)piperazine-N′-(4-butanesulfonic acid)),TAPS(N-tris(hydroxymethyl)methyl-3-aminopropanesulfonic acid), AMPD(2-amino-2-methyl-1,3-propanediol), and/or any other buffers known tothose of skill in the art.

A complexation-enhancing agent can be added to a formulation of theinvention. When such an agent is present, the ratio ofcyclodextrin/active agent can be changed. A complexation-enhancing agentis a compound, or compounds, that enhance(s) the complexation of theactive agent with the cyclodextrin. Suitable complexation enhancingagents include one or more pharmacologically inert water solublepolymers, hydroxy acids, and other organic compounds typically used inliquid formulations to enhance the complexation of a particular agentwith cyclodextrins.

Hydrophilic polymers can be used as complexation-enhancing,solubility-enhancing and/or water activity reducing agents to improvethe performance of formulations containing a cyclodextrin. Loftsson hasdisclosed a number of polymers suitable for combined use with acyclodextrin (underivatized or derivatized) to enhance the performanceand/or properties of the cyclodextrin. Suitable polymers are disclosedin Pharmazie (2001), 56(9), 746-747; International Journal ofPharmaceutics (2001), 212(1), 29-40; Cyclodextrin: From Basic Researchto Market, International Cyclodextrin Symposium, 10th, Ann Arbor, Mich.,United States, May 21-24, 2000 (2000), 10-15 (Wacker Biochem Corp.;Adrian, Mich.); PCT International Publication No. WO 9942111; Pharmazie,53(11), 733-740 (1998); Pharm. Technol. Eur., 9(5), 26-34 (1997); J.Pharm. Sci. 85(10), 1017-1025 (1996); European Patent ApplicationEP0579435; Proceedings of the International Symposium on Cyclodextrins,9th, Santiago de Comostela, Spain, May 31-Jun. 3, 1998 (1999), 261-264(Editor(s): Labandeira, J. J. Torres; Vila-Jato, J. L. Kluwer AcademicPublishers, Dordrecht, Neth); S.T.P. Pharma Sciences (1999), 9(3),237-242; ACS Symposium Series (1999), 737(Polysaccharide Applications),24-45; Pharmaceutical Research (1998), 15(11), 1696-1701; DrugDevelopment and Industrial Pharmacy (1998), 24(4), 365-370;International Journal of Pharmaceutics (1998), 163(1-2), 115-121; Bookof Abstracts, 216th ACS National Meeting, Boston, Aug. 23-27 (1998),CELL-016, American Chemical Society; Journal of Controlled Release,(1997), 44/1 (95-99); Pharm. Res. (1997) 14(11), S203; InvestigativeOphthalmology & Visual Science, (1996), 37(6), 1199-1203; Proceedings ofthe International Symposium on Controlled Release of Bioactive Materials(1996), 23rd, 453-454; Drug Development and Industrial Pharmacy (1996),22(5), 401-405; Proceedings of the International Symposium onCyclodextrins, 8th, Budapest, Mar. 31-Apr. 2, (1996), 373-376.(Editor(s): Szejtli, I.; Szente, L. Kluwer: Dordrecht, Neth.);Pharmaceutical Sciences (1996), 2(6), 277-279; European Journal ofPharmaceutical Sciences, (1996) 4(SUPPL.), S144; Third European Congressof Pharmaceutical Sciences Edinburgh, Scotland, UK Sep. 15-17, 1996;Pharmazie, (1996), 51(1), 39-42; Eur. J. Pharm. Sci. (1996), 4(Suppl.),S143; U.S. Pat. No. 5,472,954 and U.S. Pat. No. 5,324,718; InternationalJournal of Pharmaceutics (Netherlands), (Dec. 29, 1995) 126, 73-78;Abstracts of Papers of the American Chemical Society, (2 Apr. 1995)209(1), 33-CELL; European Journal of Pharmaceutical Sciences, (1994) 2,297-301; Pharmaceutical Research (New York), (1994) 11(10), S225;International Journal of Pharmaceutics (Netherlands), (Apr. 11, 1994)104, 181-184; and International Journal of Pharmaceutics (1994), 110(2),169-77, the entire disclosures of which are hereby incorporated byreference.

Other suitable polymers are well-known excipients commonly used in thefield of pharmaceutical formulations and are included in, for example,Remington's Pharmaceutical Sciences, 18th Edition, Alfonso R. Gennaro(editor), Mack Publishing Company, Easton, Pa., 1990, pp. 291-294;Alfred Martin, James Swarbrick and Arthur Commarata, Physical Pharmacy.Physical Chemical Principles in Pharmaceutical Sciences, 3rd edition(Lea & Febinger, Philadelphia, Pa., 1983, pp. 592-638); A. T. Florenceand D. Altwood, (Physicochemical Principles of Pharmacy, 2nd Edition,MacMillan Press, London, 1988, pp. 281-334. The entire disclosures ofthe references cited herein are hereby incorporated by references. Stillother suitable polymers include water-soluble natural polymers,water-soluble semi-synthetic polymers (such as the water-solublederivatives of cellulose) and water-soluble synthetic polymers. Thenatural polymers include polysaccharides such as inulin, pectin, alginderivatives (e.g. sodium alginate) and agar, and polypeptides such ascasein and gelatin. The semi-synthetic polymers include cellulosederivatives such as methylcellulose, hydroxyethylcellulose,hydroxypropyl cellulose, their mixed ethers such as hydroxypropylmethylcellulose and other mixed ethers such as hydroxyethylethylcellulose and hydroxypropyl ethylcellulose, hydroxypropylmethylcellulose phthalate and carboxymethylcellulose and its salts,especially sodium carboxymethylcellulose. The synthetic polymers includepolyoxyethylene derivatives (polyethylene glycols) and polyvinylderivatives (polyvinyl alcohol, polyvinylpyrrolidone and polystyrenesulfonate) and various copolymers of acrylic acid (e.g. carbomer). Othernatural, semi-synthetic and synthetic polymers not named here which meetthe criteria of water solubility, pharmaceutical acceptability andpharmacological inactivity are likewise considered to be within theambit of the present invention.

An emulsifying agent is intended to mean a compound that aids theformation of an emulsion. An emulsifier can be used to wet thecorticosteroid and make it more amenable to dissolution. Emulsifiers foruse herein include, but are not limited to, polyoxyetheylene sorbitanfatty esters or polysorbates, including, but not limited to,polyethylene sorbitan monooleate (Polysorbate 80), polysorbate 20(polyoxyethylene (20) sorbitan monolaurate), polysorbate 65(polyoxyethylene (20) sorbitan tristearate), polyoxyethylene (20)sorbitan mono-oleate, polyoxyethylene (20) sorbitan monopalmitate,polyoxyethylene (20) sorbitan monostearate; lecithins; alginic acid;sodium alginate; potassium alginate; ammonium alginate; calciumalginate; propane-1,2-diol alginate; agar; carrageenan; locust bean gum;guar gum; tragacanth; acacia; xanthan gum; karaya gum; pectin; amidatedpectin; ammonium phosphatides; microcrystalline cellulose;methylcellulose; hydroxypropylcellulose; hydroxypropylmethylcellulose;ethylmethylcellulose; carboxymethylcellulose; sodium, potassium andcalcium salts of fatty acids; mono- and di-glycerides of fatty acids;acetic acid esters of mono- and di-glycerides of fatty acids; lacticacid esters of mono- and di-glycerides of fatty acids; citric acidesters of mono- and di-glycerides of fatty acids; tartaric acid estersof mono- and di-glycerides of fatty acids; mono- and diacetyltartaricacid esters of mono- and di-glycerides of fatty acids; mixed acetic andtartaric acid esters of mono- and di-glycerides of fatty acids; sucroseesters of fatty acids; sucroglycerides; polyglycerol esters of fattyacids; polyglycerol esters of polycondensed fatty acids of castor oil;propane-1,2-diol esters of fatty acids; sodium stearoyl-2-lactylate;calcium stearoyl-2-lactylate; stearoyl tartrate; sorbitan monostearate;sorbitan tristearate; sorbitan monolaurate; sorbitan monooleate;sorbitan monopalmitate; extract of quillaia; polyglycerol esters ofdimerised fatty acids of soya bean oil; oxidatively polymerised soyabean oil; and pectin extract.

As used herein, the term “stabilizer” is intended to mean a compoundused to stabilize the therapeutic agent against physical, chemical, orbiochemical process that would reduce the therapeutic activity of theagent. Suitable stabilizers include, by way of example and withoutlimitation, albumin, sialic acid, creatinine, glycine and other aminoacids, niacinamide, sodium acetyltryptophonate, zinc oxide, sucrose,glucose, lactose, sorbitol, mannitol, glycerol, polyethylene glycols,sodium caprylate and sodium saccharin and other known to those ofordinary skill in the art.

As used herein, the term “tonicity modifier” is intended to mean acompound or compounds that can be used to adjust the tonicity of theliquid formulation. Suitable tonicity modifiers include glycerin,lactose, mannitol, dextrose, sodium chloride, sodium sulfate, sorbitol,trehalose and others known to those of ordinary skill in the art. Othertonicity modifiers include both inorganic and organic tonicity adjustingagents. Tonicity modifiers include, but are not limited to, ammoniumcarbonate, ammonium chloride, ammonium lactate, ammonium nitrate,ammonium phosphate, ammonium sulfate, ascorbic acid, bismuth sodiumtartrate, boric acid, calcium chloride, calcium disodium edetate,calcium gluconate, calcium lactate, citric acid, dextrose,diethanolamine, dimethylsulfoxide, edetate disodium, edetate trisodiummonohydrate, fluorescein sodium, fructose, galactose, glycerin, lacticacid, lactose, magnesium chloride, magnesium sulfate, mannitol,polyethylene glycol, potassium acetate, potassium chlorate, potassiumchloride, potassium iodide, potassium nitrate, potassium phosphate,potassium sulfate, proplyene glycol, silver nitrate, sodium acetate,sodium bicarbonate, sodium biphosphate, sodium bisulfate, sodium borate,sodium bromide, sodium cacodylate, sodium carbonate, sodium chloride,sodium citrate, sodium iodide, sodium lactate, sodium metabisulfite,sodium nitrate, sodium nitrite, sodium phosphate, sodium propionate,sodium succinate, sodium sulfate, sodium sulfite, sodium tartrate,sodium thiosulfate, sorbitol, sucrose, tartaric acid, triethanolamine,urea, urethan, uridine and zinc sulfate. In one embodiment, the tonicityof the liquid formulation approximates the tonicity of the tissues inthe respiratory tract.

An osmotic agent can be used in the compositions to enhance the overallcomfort to the patient upon delivery of the corticosteroid composition.Osmotic agents can be added to adjust the tonicity of SAE-CD containingsolutions. Osmolality is related to concentration of SAE-CD in water. AtSBE7-β-CD concentrations below about 11-13% w/v, the solutions arehypotonic or hypoosmotic with respect to blood and at SBE7-β-CDconcentrations above about 11-13% w/v the SBE7-β-CD containing solutionsare hypertonic or hyperosmotic with respect to blood. When red bloodcells are exposed to solutions that are hypo- or hypertonic, they canshrink or swell in size, which can lead to hemolysis. As noted above andin FIG. 1, SBE-CD is less prone to induce hemolysis than otherderivatized cyclodextrins. Suitable osmotic agents include any lowmolecular weight water-soluble species pharmaceutically approved forpulmonary and nasal delivery such as sodium chloride, lactose andglucose. The formulation of the invention can also include biologicalsalt(s), potassium chloride, or other electrolyte(s).

As used herein, the term “antifoaming agent” is intended to mean acompound or compounds that prevents or reduces the amount of foamingthat forms on the surface of the liquid formulation. Suitableantifoaming agents include dimethicone, simethicone, octoxynol, ethanoland others known to those of ordinary skill in the art.

As used herein, the term “bulking agent” is intended to mean a compoundused to add bulk to the lyophilized product and/or assist in the controlof the properties of the formulation during lyophilization. Suchcompounds include, by way of example and without limitation, dextran,trehalose, sucrose, polyvinylpyrrolidone, lactose, inositol, sorbitol,dimethylsulfoxide, glycerol, albumin, calcium lactobionate, and othersknown to those of ordinary skill in the art.

As used herein, the term “cryoprotectant” is intended to mean a compoundused to protect an active therapeutic agent from physical or chemicaldegradation during lyophilization. Such compounds include, by way ofexample and without limitation, dimethyl sulfoxide, glycerol, trehalose,propylene glycol, polyethylene glycol, and others known to those ofordinary skill in the art.

A solubility-enhancing agent or solubility enhancer can be added to theformulation of the invention. A solubility-enhancing agent is acompound, or compounds, that enhance(s) the solubility of thecorticosteroid when in an aqueous liquid carrier. When anothersolubility enhancing agent is present, the ratio of SAE-CD tocorticosteroid can be changed, thereby reducing the amount of SAE-CDrequired to dissolve the corticosteroid. Suitable solubility enhancingagents include one or more cyclodextrins, cyclodextrin derivatives,SAE-CD, organic solvents, detergents, soaps, surfactant and otherorganic compounds typically used in parenteral formulations to enhancethe solubility of a particular agent. Exemplary solubility enhancers aredisclosed in U.S. Pat. No. 6,451,339; however, other surfactants used inthe pharmaceutical industry can be used in the formulation of theinvention. Some suitable cyclodextrin include underivatizedcyclodextrins and cyclodextrin derivatives, such as SAE-CD, SAE-CDderivatives, hydroxyalkyl ether cyclodextrin and derivatives, alkylether cyclodextrin and derivatives, sulfated cyclodextrin andderivatives, hydroxypropyl-β-cyclodextrin, 2-HP-β-CD,methyl-β-cyclodextrin, carboxyalkyl thioether derivatives, succinylcyclodextrin and derivatives, and other cyclodextrin suitable forpharmaceutical use. SAE-CD cyclodextrins are particularly advantageous.

Suitable organic solvents that can be used in the formulation include,for example, ethanol, glycerin, poly(ethylene glycol), propylene glycol,poloxamer, aqueous forms thereof and others known to those of ordinaryskill in the art.

It should be understood that compounds used in the art of pharmaceuticalformulations generally serve a variety of functions or purposes. Thus,if a compound named herein is mentioned only once or is used to definemore than one term herein, its purpose or function should not beconstrued as being limited solely to that named purpose(s) orfunction(s).

An active agent contained within the present formulation can be presentas its pharmaceutically acceptable salt. As used herein,“pharmaceutically acceptable salt” refers to derivatives of thedisclosed compounds wherein the active agent is modified by reacting itwith an acid or base as needed to form an ionically bound pair. Examplesof pharmaceutically acceptable salts include conventional non-toxicsalts or the quaternary ammonium salts of the parent compound formed,for example, from non-toxic inorganic or organic acids. Suitablenon-toxic salts include those derived from inorganic acids such ashydrochloric, hydrobromic, sulfuric, sulfonic, sulfamic, phosphoric,nitric and others known to those of ordinary skill in the art. The saltsprepared from organic acids such as amino acids, acetic, propionic,succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic,pamoic, maleic, hydroxymaleic, phenylacetic, glutamic, benzoic,salicylic, sulfanilic, 2-acetoxybenzoic, fumaric, toluenesulfonic,methanesulfonic, ethane disulfonic, oxalic, isethionic, and others knownto those of ordinary skill in the art. The pharmaceutically acceptablesalts of the present invention can be synthesized from the parent activeagent which contains a basic or acidic moiety by conventional chemicalmethods. Lists of other suitable salts are found in Remington'sPharmaceutical Sciences, 17^(th), ed., Mack Publishing Company, Easton,Pa., 1985, the relevant disclosure of which is hereby incorporated byreference.

The phrase “pharmaceutically acceptable” is employed herein to refer tothose compounds, materials, compositions, and/or dosage forms which are,within the scope of sound medical judgment, suitable for use in contactwith the tissues of human beings and animals without excessive toxicity,irritation, allergic response, or other problem or complication,commensurate with a reasonable benefit/risk ratio.

As used herein, the term “patient” or “subject” are taken to mean warmblooded animals such as mammals, for example, cats, dogs, mice, guineapigs, horses, bovine cows, sheep and humans.

A formulation of the invention will comprise an active agent present inan effective amount, effective dose or therapeutically effective dose.By the term “effective amount” or “effective dose” or “therapeuticallyeffective dose”, is meant the amount or quantity of active agent that issufficient to elicit the required or desired response, or in otherwords, the amount that is sufficient to elicit an appreciable biologicalresponse when administered to a subject.

In view of the above description and the examples below, one of ordinaryskill in the art will be able to practice the invention as claimedwithout undue experimentation. The foregoing will be better understoodwith reference to the following examples that detail certain proceduresfor the preparation of formulations according to the present invention.All references made to these examples are for the purposes ofillustration. The following examples should not be consideredexhaustive, but merely illustrative of only a few of the manyembodiments contemplated by the present invention.

Example 1

Exemplary formulations according to the invention were made according tothe following general procedures.

Method A

Cyclodextrin is dissolved in water (or buffer) to form a solutioncontaining a known concentration of cyclodextrin. This solution is mixedwith an active agent in solid, suspension, gel, liquid, paste, powder orother form while mixing, optionally while heating to form an inhalablesolution.

Method B

A known amount of substantially dry cyclodextrin is mixed with a knownamount of substantially dry active agent. A liquid is added to themixture to form a suspension, gel, solution, syrup or paste whilemixing, optionally while heating and optionally in the presence of oneor more other excipients, to form an inhalable solution.

Method C

A known amount of substantially dry cyclodextrin is added to asuspension, gel, solution, syrup or paste comprising a known amount ofactive agent while mixing, optionally while heating and optionally inthe presence of one or more other excipients, to form an inhalablesolution.

The methods of this example may be modified by the inclusion of awetting agent in the composition in order to facilitate dissolution andsubsequent inclusion complexation of the corticosteroid. A surfactant,soap, detergent or emulsifying agent can be used as a wetting agent.

Method D

To a solution comprising a known concentration or amount of SAE-CD,aqueous liquid carrier, and optionally one or more other excipients, isadded a molar excess of the corticosteroid based upon the molar ratio ofSAE-CD to corticosteroid at the point of saturated solubility of thecorticosteroid, in the presence of the SAE-CD, as determined herein. Forexample, corticosteroid would be added at a 5%, 10%, 15%, 20%, 25%, 30%or greater molar excess. The components are mixed until equilibration,the point at which there is only a minor change in the concentration ofbudesonide over a one-hour period of time. Then, the excesscorticosteroid is removed leaving behind the target solution of theinvention.

The budesonide is added to the SAE-CD-containing solution as either asolid or suspension in an aqueous liquid carrier, which can be water,buffer, aqueous alcohol, aqueous organic solvent or a combinationthereof. The alcohol and organic solvent are of a pharmaceuticallyacceptable grade, such as ethanol, propylene glycol, and others asdescribed herein.

Method E

The SAE-CD and corticosteroid are triturated to form a mixture. Then, anaqueous liquid carrier is added to the mixture form the target solutionof the invention.

The trituration can be conducted dry or in the presence of moisture,water, buffer, alcohol, surfactant, organic solvent, glycerin,poly(ethylene glycol), poloxamer, or a combination thereof.

Method F

Any of the methods herein are conducted in the presence of heat, e.g. ata temperature of least 40° C.

Method G

Any of the methods herein are conducted with cooling, e.g. at atemperature of less than 20° C. or less than 10° C. or less than 5° C.

Method H

Any of the methods herein are conducted in the presence of high shearmixing such as with a sonicator, narrow gauge syringe(s),mixer/homogenizer (POLYTRON from KINEMATICA, Europe; FLUKO, Shanghai,China; ULTIMAGRAL from GEA Niro, Inc., Columbia, Md.), rotor-statormixer, or saw tooth mixer.

Method I

Any of the methods herein are conducted under reduced pressure.

Example 2

The MMD of nebulized solutions containing SBE7-β-CD and budesonide wasdetermined as follows.

Placebo solutions of three different cyclodextrins were prepared atdifferent concentrations. Two ml of the solutions were added to the cupof a Pari LC Plus nebulizer supplied with air from a Pari Proneb Ultracompressor. The particle size of the emitted droplets was determinedusing a Malvern Mastersizer S laser light scattering instrument.

Example 3

The stability of liquid formulations containing SAE-CD was determined byHPLC chromatography of aliquots periodically drawn from the liquid instorage.

Citrate-phosphate (McIlvaines) buffer solutions at a pH of 4, 5, 6, 7,or 8 were prepared by mixing various portions of 0.01M citric acid with0.02 M Na₂HPO₄. These stock solutions contained 5% w/w Captisol.Approximately 250 μg/mL of budesonide was dissolved in each buffersolution. Aliquots of the solutions were stored at 40° C., 50° C. and60° C. Control samples were stored at 5° C. but are not reported here.HPLC analysis of the samples was performed initially and after 1, 2, and3 months storage.

The HPLC conditions included:

Instrument: PE Series 200 Column: Phenomenex Luna C18(2) 4.6 × 150 mm 3um Mobile Phase: 58% Phosphate Buffer pH 3.4/39.5% ACN/2.5% MeOH MobilePhase 100% A (isocratic) Program: Wavelength 240 Flow Rate: 6.6 mL/minStandard Range: Seven standards—1 to 500 μg/mL

Example 4

The viscosity of aqueous solutions containing SAE-CD were measured usinga cone and plate viscometer.

A Brookfield Programmable DV-III+ Rheometer, CPE-40 cone and CPE 40Yplate (Brookfield Engineering Laboratories, Middleboro, Mass.) was usedto make measurements on 0.5 ml samples at 1, 2, 3, 5 and 10 rpm, Sampleswere sheered for approximately 5 revolutions prior to each measurement.This allowed accurate theological characterization of the samples. Thetemperature of all samples was equilibrated to 25+/−1 degree centigradeusing a double wall viscometer cone supplied with water from anelectronically controlled thermostatic circulating water bath (Model,8001, Fisher Scientific, Pittsburgh, Pa.) The viscometer was calibratedusing 5 and 50 centipoise using silicon oil calibration standards.Viscosity measurements were made at 5 or more rotation speeds to lookfor sheer thinning behavior (viscosities that decrease as the rate ofsheer increases). Higher rotation speeds result in increased rates ofsheer.

Example 5

Nebulizer output rate as a function of SAE-CD concentration was measuredaccording to the following general procedure.

Nebulizer Output was tested using Pari LC Plus Nebulizer with a PariProNeb Ultra Air Compressor (Minimum Nebulizer Volume=2 ml, MaximumNebulizer Volume=8 ml) for solutions containing 43%, 21.5%, 10.75% and5.15% w/w SBE7-β-CD. Percentage of sample emitted was estimatedgravimetrically. The nebulizer cup was weighed before and afternebulization was complete. Nebulization Time was defined as the durationof time when nebulizer run was started until the time of first sputter.Nebulizer Output Rate was calculated by dividing % Emitted withNebulization Time.

Example 6

Preparation of an inhalable solution containing budesonide.

A buffer solution containing 3 mM Citrate Buffer and 82 mM NaCl at pH4.45 is prepared. ˜12.5 grams of CAPTISOL was placed into a 250 mlvolumetric flask. ˜62.5 mg of budesonide was placed into the same flask.Flask was made to volume with the 3 mM citrate buffer/82 mM NaClsolution. The flask was well-mixed on a vortexer for 10 minutes andsonicated for 10 minutes. The flask was stirred over weekend withmagnetic stirrer. Stirring was stopped after ˜62 hours and flask wasrevortexed and resonicated again for 10 minutes each. The solution wasfiltered through a 0.22 μm Durapore Millex-GV Millipore syringe filterunit. The first few drops were discarded before filter rest of solutioninto an amber glass jar with a Teflon-lined screw cap. Sampleconcentration was ˜237 μg/ml.

Example 7

Preparation of an inhalable solution containing budesonide.

Approximately 5 grams of CAPTISOL was placed into a 100 mL volumetricflask. ˜26.3 mg of budesonide was placed into the same flask. The flaskwas made to volume with the 3 mM citrate buffer/82 mM NaCl solution. Themixture was well-mixed on a vortexer for 10 minutes and sonicated for 10minutes. The mixture was stirred overnight with a magnetic stirrer.Stirring was stopped after ˜16 hours and flask was revortexed andresonicated again for 10 minutes each. The solution was filtered through0.22 μm Durapore Millex-GV Millipore syringe filter unit. The first 5drops were discarded before filter rest of solution into an amber glassjar with a Teflon-lined screw cap. Sample was analyzed to be 233 μgbudesonide/ml.

Example 8

Preparation of an inhalable solution containing budesonide.

The procedure of Example 7 was followed except that 12.5 g of CAPTISOL,62.5 mg of budesonide and about 250 ml of buffer were used. Sufficientdisodium EDTA was added to prepare a solution having an EDTAconcentration of about 0.01 or 0.05% wt/v EDTA.

Example 9

Preparation of a solution containing SAE-CD and budesonide as preparedfrom a PULMICORT RESPULES suspension.

Method A

To the contents of one or more containers of the Pulmicort Respules(nominally 2 mL of the suspension), about 50 mg (corrected for watercontent) of CAPTISOL was added per mL of Respule and mixed or shakenwell for several minutes. After standing from about 30 minutes toseveral hours, the solution was used as is for in vitrocharacterization. In addition to budesonide and water, the PULMICORTRESPULE (suspension) also contains the following inactive ingredientsper the label: citric acid, sodium citrate, sodium chloride, disodiumEDTA and polysorbate 80.

Method B

Weigh approximately 200 mg amounts of CAPTISOL (corrected for watercontent) into 2-dram amber vials. Into each vial containing the weighedamount of CAPTISOL empty the contents of two Pulmicort Respulescontainers (0.5 mg/2 mL, Lot #308016 Feb05) by gently squeezing thedeformable plastic container to the last possible drop. The Respuleswere previously swirled to re-suspend the budesonide particles. Thevials are screw capped, mixed vigorously by vortex and then foilwrapped. The material can be kept refrigerated until use.

The inhalable liquid composition prepared according to any of thesemethods can be used in any known nebulizer. By converting the suspensionto a liquid, an improvement in delivery of budesonide (a corticosteroid)is observed.

Example 10

Other solutions according to the invention can be prepared as detailedbelow.

Mg per ml (as prepared) Mg per ml Concentrate Concentrate (per target)Component A B Final Solution Budesonide EP 1 ~1.6 (sat'd) 0.250 CAPTISOL200 200 50 Sodium Citrate 0 0 0.44 tribasic dihydrate Citric Acid 0 00.32 Sodium Chloride 0 0 4.8 Disodium EDTA 0 0 0-0.5 Polysorbate 80 0 00-1   (Tween 80) Water Qs Qs QS with buffer containing CAPTISOL orbudesonide

-   -   Dilute Concentrate A at a ratio of 1 to 4 with pH 4.5 salinated        citrate buffer (4 mM containing 109 mM sodium chloride) to        contain 5% w/v CAPTISOL on an anhydrous basis. Filter the        diluted concentrate through a 0.22 μm Millipore Durapore        Millex-GV syringe filter unit. Assay the filtered solution by        HPLC then add supplemental budesonide as needed to give a        solution final concentration of about 250 μg/mL (±<5%).    -   Dilute Concentrate B at a ratio of 1 to 4 with pH 4.5 salinated        citrate buffer (4 mM containing 109 mM sodium chloride) to        contain 5% w/v CAPTISOL on an anhydrous basis. Filter the        diluted concentrate through a 0.22 μm Millipore Durapore        Millex-GV syringe filter unit. Assay the filtered solution by        HPLC then dilute further with pH 4.5 salinated citrate buffer (3        mM containing 82 mM sodium chloride containing 5% w/v CAPTISOL)        as required to give a final solution concentration of about 250        μg/mL (±<5%). This technique takes advantage of the excess solid        budesonide used to saturate the solution.

Example 11

Clarity of solutions was determined by visual inspection orinstrumentally. A clear solution is at least clear by visual inspectionwith the unaided eye.

Example 12

The following method was used to determine the performance ofnebulization compositions emitted from a nebulizer according to FIGS. 10a-10 b.

Two ml of the test CD solution or Pulmicort suspension was accuratelypipetted by volumetric pipettes into a clean nebulizer cup prior tostarting each experiment. The test nebulizer was assembled and chargedwith the test inhalation solution or suspension according to themanufacturer instructions. The end of the mouthpiece was placed at aheight of approximately 18 cm from the platform of the MALVERNMASTERSIZER to the middle point of tip of the nebulizer mouthpiece. Avacuum source was positioned opposite the mouthpiece approximately 6 cmaway to scavenge aerosol after sizing. The distance between themouthpiece and the detector was approximately 8 cm. The center of themouthpiece was level with the laser beam (or adjusted as appropriate,depending on the individual design of each nebulizer). The laser passedthrough the center of the emitted cloud when the nebulizer was running.Measurements were manually started 15 seconds into nebulization. Datacollection started when beam obscuration reached 10% and was averagedover 15,000 sweeps (30 seconds). Scattered light intensity data on thedetector rings was modeled using the “Standard-Wet” model. Channels 1and 2 were killed due to low relative humidity during measurement toprevent beam steering. The volume diameter of droplets defining 10, 50(volume median), and 90% of the cumulative volume undersize wasdetermined. (Dv10 is the size below which 10% of the volume of materialexists, Dv50 is the size below which 50% of the volume of materialexists and Dv90 is the size below which 90% of the volume of materialexists.

The procedure above may be practiced with slight modification on aMALVERN SPRAYTEC to determine the particle size of droplets emitted by anebulizer.

Example 13

Solutions of budesonide with and without SBE7-β-CD were prepared at twodifferent pHs (4 and 6) and stored at 2 different temperatures (60° C.and 80° C.). Citrate buffers (50 mM) at each pH value were prepared bymixing differing portions of 50 mM citric acid and 50 mM sodium citrate(tribasic, dihydrate) solutions. To achieve a concentration ofbudesonide in the buffers without SBE7-β-CD sufficient for accuratemeasurement, the budesonide was dissolved first in 100% ethyl alcohol.An aliquot of the ethanol/budesonide solution was then added drop-wisewith stirring to each buffer solution. The theoretical budesonideconcentration was 100 μg/mL with a final ethanolic content of 5% in eachbuffer. All solution preps and procedures involving budesonide were donein a darkened room under red light. After shaking solutions for 24hours, both buffer solutions were filtered through Millipore Millex-GV0.22 μm syringe filters to remove any solid that had precipitated (nosignificant amounts observed) from the solutions. The final budesonideconcentration was about 50 μg/mL. Both the pH 4 and 6 solutions weresplit in two, and solid SBE7-β-CD was added to one of the portions tocreate solutions with and without 1% w/v SBE7-β-CD at each pH. Eachsolution was aliquoted into individual amber vials. They were thenplaced in ovens at 60° C. and 80° C. Sample vials were removed from theovens and analyzed by HPLC at 0, 96, 164, and 288 hours. The HPLC assayconditions are summarized below.

Chromatographic Conditions (Adapted from Hou, S., Hindle, M., and Byron,P. R. A. Stability- Indicating HPLC Assay Method for Budesonide. Journalof Pharmaceutical and Biomedical Analysis, 2001; 24: 371-380.)Instrument: PE Series 200 Column: Phenomenex Luna C18(2) 4.6 × 150 mm 3um Mobile Phase: 58% Phosphate Buffer pH 3.4/39.5% ACN/2.5% MeOH MobilePhase 100% A (isocratic) Program: Wavelength 240 nm Flow Rate: 0.6mL/min Standard Range: Seven standards—1 to 500 μg/mL

Example 14

Preparation and use of a combination solution containing SAE-CD,budesonide, and albuterol sulfate.

A budesonide solution is prepared per EXAMPLE 9 (mixing SAE-CD with thePULMICORT RESPULES suspension) and added to 3 ml of a solutioncontaining 2.5 mg albuterol (The World Health Organization recommendedname for albuterol base is salbutamol) provided as albuterol sulfate.The albuterol solution is commercially available prediluted and soldunder the name PROVENTIL® Inhalation Solution, 0.083%, or prepared froma commercially available concentrate 0.5% (sold under the names:PROVENTIL® Solution for inhalation and VENTOLIN® Inhalation Solution).

To provide the required dose for children 2 to 12 years of age, theinitial dosing should be based upon body weight (0.1 to 0.15 mg/kg perdose), with subsequent dosing titrated to achieve the desired clinicalresponse. Dosing should not exceed 2.5 mg three to four times daily bynebulization. The appropriate volume of the 0.5% inhalation solutionshould be diluted in sterile normal saline solution to a total volume of3 mL prior to administration via nebulization. To provide 2.5 mg, 0.5 mLof the concentrate is diluted to 3 mL with sterile normal saline. Thealbuterol aqueous solutions also contain benzalkonium chloride; andsulfuric acid is used to adjust the pH to between 3 and 5.Alternatively, an aqueous solution of an appropriate strength ofalbuterol may be prepared from albuterol sulfate, USP with or withoutthe added preservative benzalkonium chloride and pH adjustment usingsulfuric acid may also be unnecessary when combining with thecorticosteroid solution. Furthermore the volume containing theappropriate dose of corticosteroid may be decreased four-fold asdescribed in the following example allowing the total volume to be lessand the time of administration to diminish accordingly.

Example 15

Preparation and use of a combination solution containing SAE-CD,budesonide, and albuterol sulfate or levalbuterol HCl (XOPENEX).

A citrate buffer (3 mM pH 4.5) was prepared as follows. Approximately62.5 mg of citric acid was dissolved in and brought to volume with waterin one 100 ml volumetric flask. Approximately 87.7 mg of sodium citratewas dissolved in and brought to volume with water in another 100 mLvolumetric flask. In a beaker the sodium citrate solution was added tothe citric acid solution until the pH was approximately 4.5.

Approximately 10.4 mg of budesonide and 1247.4 mg of Captisol wereground together with a mortar and pestle and transferred to a 10 mLflask. Buffer solution was added, and the mixture was vortexed,sonicated and an additional 1.4 mg budesonide added. After shakingovernight, the solution was filtered through a 0.22 μm DuraporeMillex-GV Millipore syringe filter unit. The resulting budesonideconcentration was ˜1 mg/ml. Approximately 0.5 ml of the budesonidesolution was added to a unit dose of either Proventil (2.5 mg/3 mL) orXopenex (1.25 mg/3 mL) thereby forming the combination clear liquiddosage form of the invention. The resulting mixture remained essentiallyclear for a period of at least 17 days at ambient room conditionsprotected from light.

Example 16

Preparation and use of a combination solution containing SAE-CD,budesonide, and formoterol (FORADIL® (formoterol fumarate inhalationpowder)).

The contents of one capsule containing 12 mcg of formoterol fumarateblended with 25 mg of lactose was emptied into a vial to which was added3-mL of 3 mM citrate buffer (pH 4.5) prepared as described in theprevious example. The contents of the vial were vortexed to dissolve thesolids present. The budesonide concentrate was prepared as described inthe previous example to provide a concentration of ˜1 mg/mL.Approximately 1 ml of the budesonide solution was added to theformoterol fumarate buffered solution. The resulting solution remainedessentially clear for a period of at least one month at room ambientconditions protected from light.

Example 17

Clinical evaluation of a dosage form according to the invention wasconducted by performing gamma scintigraphy analyses on subjects beforeand after administration of the dosage form by nebulization.

A single centre, four-way crossover gamma scintigraphy study to comparepulmonary delivery of budesonide via Pulmicort Respules®, andCaptisol-Enabled® budesonide formulations using a Pari LC air-jetnebulizer was conducted. The purpose of the study was to determine, bygamma scintigraphy, the intra-pulmonary deposition of radiolabeledbudesonide following nebulization of a budesonide suspension (PulmicortRespules®, Astra Zeneca, reference formulation) and a Captisol®-Enabledbudesonide solution (test formulation) in healthy male volunteers.During the course of the study, blood was collected periodically up to24 hours, and the plasma was isolated. The concentration of budesonidein the plasma was determined using a validated HPLC MS-MS method inorder to determine pharmacokinetic parameters. Dosing was conductedusing a Pari LC Plus air-jet nebulizer. The use of gamma scintigraphy inconjunction with radiolabeled study drug and/or vehicle is the standardtechnique for the quantitative assessment of pulmonary deposition andclearance of inhaled drugs and/or vehicle.

The study dosage forms consisted of: 1) 1 mg Budesonide as 2 mL×0.5mg/mL Pulmicort Respules®; or 2) 1 mg Budesonide as 2 mL×0.5 mg/mLPulmicort Respules to which 7.5% w/v Captisol® has been added.

Each subject received each of four study administrations of radiolabeledBudesonide in a non-randomized manner. A non-randomized design wasutilized for this study since the reference formulation (PulmicortRespule) must be administered first to all subjects in order todetermine the time to sputter (TTS). The TTS differed between subjects.For subsequent administrations the dose administered was controlled bythe length of administration, expressed as a fraction of the time tosputter determined following administration of the reference formulation(i.e. 25% TTS, 50% TTS and 75% TTS). It was expected that even thoughthe same concentration of budesonide would be nebulized for a shortertime, the amount of drug reaching the volunteers lungs would beessentially the same as the reference suspension for one of the legs ofthe study. Scintigraphic images were acquired using a gamma cameraimmediately after completion of dosing the volunteers.

Comparison of the image from the reference product and the 25% TTS legindicated that a greater percentage of the budesonide from the Respulewas in the stomach and throat immediately after administration. Thus agreater percentage of the budesonide reached the target lung tissue whenCaptisol was used to dissolve the budesonide. This could reduceundesirable side effects caused by the drug. One aspect of the methodand dosage form of the invention thus provides an improved method ofadministering a corticosteroid suspension-based unit dose, the methodcomprising the step of adding a sufficient amount of SAE-CD to convertthe suspension to a clear solution and then administering the clearsolution to a subject. As a result, the method of the invention providesincreased rate of administration as well as increased total pulmonarydelivery of the corticosteroid as compared to the initial unit dosesuspension formulation.

Example 18

Comparative evaluation of various forms of SAE-CD in the solubilizationof corticosteroid derivatives.

The solubility of beclomethasone dipropionate (BDP), beclomethasone17-monopropionate (B17P), beclomethasone 21-monopropionate (B21P) andbeclomethasone (unesterifed) in solutions containing CAPTISOL andvarious SBE_(n)γ-CD was evaluated. BDP, B17P and B21P were obtained fromHovione. Beclomethasone was obtained from Spectrum Chemicals. CAPTISOL,SBE(3.4) γ-CD, SBE(5.23) γ-CD and SBE(6.1) γ-CD were provided by CyDex,Inc. (Lenexa, Kans.). γ-CD was obtained from Wacker Chemical Co.SBE(5.24) γ-CD and SBE(7.5) γ-CD were provided by the University ofKansas.

A 0.04M solution of each selected CD was prepared. Each form ofbeclomethasone required 2 ml of CD solution, therefore the 0.04Msolutions were prepared in 20 or 25 mL volumetric flasks in duplicate(N=2). The following table indicates the amount of each CD used afteraccounting for the content of water in each CD.

CD MW (g/mole) mg of CD (volume) SBE(6.7) β-CD 2194.6 2297.0 (25 ml)γ-CD 1297 1433.0 (25 ml) SBE(3.4) γ-CD 1834.9 1891.6 (25 ml) SBE(5.24)γ-CD 2119.5 1745.7 (20 ml) SBE(6.1) γ-CD 2261.9 1866.8 (20 ml) SBE(7.5)γ-CD 2483.3 2560.0 (25 ml)

Beclomethasone forms were weighed in amounts in excess of theanticipated solubilities directly into 2-dram Teflon-lined screw-cappedvials. These amounts typically provided approximately 6 mg/mL of solids.Each vial then received 2 ml of the appropriate CD solution. The vialswere vortexed and sonicated for about 10 minutes to aid in wetting thesolids with the fluid. The vials were then wrapped in aluminum foil toprotect from light and placed on a lab quake for equilibration. Thevials were visually inspected periodically to assure that the solidswere adequately being wetted and in contact with the fluid. The timepoints for sampling were at 24 hrs for all samples and 72 hours for BDPonly.

Solutions of SBE(6.1) γ-CD were prepared at 0.04, 0.08, and 0.1M andsolutions of SBE (5.23) γ-CD were prepared at only 0.04 and 0.08M.Beclomethasone dipropionate was weighed in amounts in excess of theanticipated solubilities directly into 2-dram teflon-lined screw-cappedvials. These amounts typically provided approximately 2 mg/mL of solids.Each vial then received 2 mL of the appropriate CD solution (N=1). Thevials were vortexed and sonicated for about 10 minutes to aid in wettingthe solids with the fluid. The vials were then wrapped in aluminum foilto protect from light and placed on a lab quake for a five-dayequilibration.

Solutions of γ-CD were prepared at 0.01 and 0.02M. Beclomethasonedipropionate was weighed in amounts in excess of the anticipatedsolubilities directly into 2-dram teflon-lined screw-capped vials. Theseamounts typically provided approximately 2 mg/mL of solids. Each vialthen received 2 mLs of the γ-CD solution (N=2). A solution was alsoprepared to measure the intrinsic solubility of BDP using HPLC gradewater in place of the CD. The samples were wrapped in foil and placed ona lab quake for five days.

At the end of the equilibration time for each stage, the vials werecentrifuged and 1 ml of the supernatant removed. The removed supernatantwas then filtered using the Durapore PVDF 0.22 μm syringe filter(discarded first few drops), and diluted with the mobile phase to anappropriate concentration within the standard curve. The samples werethen analyzed by HPLC to determine concentration of solubilizedcorticosteroid.

Example 19

Preparation and use of a combination solution containing SAE-CD,budesonide, and formoterol fumarate.

Formoterol fumarate dry powder is blended with Captisol dry powder whichare both sized appropriately to provide for content uniformity at aweight ratio of 12 mcg formoterol fumarate/100 mg Captisol. An amount ofpowder blend corresponding to a unit dose of formoterol fumarate isplaced in a suitable unit dose container such as a HPMC capsule forlater use or is added directly to a unit dose of Pulmicort Respulesbudesonide inhalation suspension (500 mcg/2 mL), then mixed to achievedissolution of all solids (a clear solution) and placed in the nebulizerreservoir for administration.

Example 20

Preparation and use of a combination solution containing SAE-CD,budesonide, and ipratropium bromide.

A budesonide solution is prepared as per EXAMPLE 9 and added to aipratropium bromide solution that is commercially available and soldunder the name ATROVENT® Inhalation Solution Unit Dose. ATROVENT®(ipratropium bromide) Inhalation Solution is 500 mcg (1 unit dose Vial)administered three to four times a day by oral nebulization, with doses6 to 8 hours apart. ATROVENT® inhalation solution unit dose Vialscontain 500 mcg ipratropium bromide anhydrous in 2.5 ml sterile,preservative-free, isotonic saline solution, pH-adjusted to 3.4 (3 to 4)with hydrochloric acid. Furthermore the volume containing theappropriate dose of corticosteroid may be decreased four-fold asdescribed in the above example (budesonide concentrate ˜1 mg/mL)allowing the total volume to be less and the time of administration todiminish accordingly.

Example 21

Evaluation of the AERONEB GO nebulizer versus a RAINDROP nebulizer witha solution comprising budesonide, aqueous liquid carrier and SAE-CD.

The AERONEB GO nebulizer (AEROGEN Inc., Mountainview, Calif.) isdetailed in U.S. Pregrant Publication No. 2005-011514 to Power et al.(application U.S. Ser. No. 10/833,932 filed Apr. 27, 2004), PCTInternational Publication No. WO 2005/009323 to Aerogen, Inc. et al.(PCT Application No. PCT/US2004/021268 filed Jul. 6, 2004), and EuropeanApplication No. EP 16426276, the entire disclosures of which are herebyincorporated by reference.

The RAINDROP nebulizer is available from Nellcor (Tyco Healthcare).

The solution of the invention used for this study was prepared accordingto Example 28.

Characterization of droplet size distribution of an aerosolized solutionusing a cascade impactor was determined according to Example 26.

Determination of total drug output and drug output rate from a nebulizercontaining a liquid of the invention was determined according to Example27.

Example 22

Evaluation of the pulsating membrane nebulizer of U.S. Pat. No.6,962,151 with a solution comprising budesonide, aqueous liquid carrierand SAE-CD. Inertial Impaction Characterization of Tc^(99m)-DTPALabelled Captisol-Enabled® Budesonide Aerosols Generated via a PariElectronic Nebulizer

The nebulizer detailed in U.S. Pat. No. 6,962,151, is also described inPCT International Application No. PCT/US00/29541 filed Oct. 27, 2000,and U.S. application Ser. No. 11/269,783 filed Nov. 7, 2005.

Aerosol characterization was conducted by standard in vitro inertialimpaction tests using an Andersen Cascade Impactor (ACI). Technetium-99m(^(99m)Tc), in the form of diethylenetriaminepenta-acetic acid (DTPA, GEHealthcare), was added to the Captisol-Enabled® Budesonide InhalationSolution (CEBUD). The suitability of ^(99m)Tc-DTPA to function as asurrogate for budesonide in CEBUD preparations was validated in thecourse of an earlier clinical trial (see Example 17). Preparation of thebudesonide solution formulation for testing was conducted as per thedescription below. It was calculated that approximately 10 MBq of^(99m)Tc should be added to the budesonide formulation (11.05 g) on theday of testing, in order to provide sufficient activity for in vitroimaging.

Two Pari electronic vibrating membrane nebulizers were used. At the coreof this electronic nebulizer is a stainless steel membrane withthousands of laser drilled holes. Laser drilling allows flexibility tocustomize particle size, ensure reproducibility, and maintain a highoutput rate with smaller particles. The perforated membrane is vibratedat high frequencies in a resonant “bending” mode which yields highparticle output rates. The nebulizer provides rapid drug delivery,efficiency, ideal particle sizing, low residual volume, and optimalperformance matched to the inhaled drug formulation (See Rajiv Dhand,Respiratory Care 2002; (12): 1406-1416). Approximately 0.5 mL of drugsolution was loaded and subsequently delivered via each nebulizer on 3separate occasions. Runs 1, 3 and 5 were conducted with Device 1 andruns 2, 4 and 6 were conducted with Device 2.

Pre-Dose

On each occasion, prior to dose delivery, the filled nebulizer wasimaged for 60 seconds on Head I of the dual head gamma camera (Axis,Philips Medical Systems). Also, the nebulizers were weighed before andafter addition of the budesonide formulation.

Inertial Impaction Testing

The nebulizer was positioned at the USP (United States Pharmacopoeia)inlet of the ACI and a flow rate of 28.3 L/min was drawn through theimpactor using a vacuum pump. Flow through the impactor was startedprior to activation of the electronic nebulizer. A stopwatch was used inorder to measure the duration of dose delivery.

The ACI test conditions were the same as those used for Pari LC Plus airjet nebulizer evaluation in the course of the earlier clinical study.

Following deposition the USP throat was removed from the ACI and imagedfor 120 seconds. The collection plates were removed from the impactorand placed on Head I of the gamma camera and imaged for 120 seconds. Theplates were subsequently washed and dried before conduct of fartherimpaction tests.

Post-Dose

On each occasion, the nebulizer weight was recorded after delivery ofthe dose. The nebulizer was imaged as described below.

Image Processing

A rectangular ROI was applied to image to the nebulizer pre-dose. ThisROI was then re-applied to image the nebulizer after dose delivery.

A rectangular ROI was also applied to the USP Inlet image.

A circular ROI was drawn around collection plate 0, copied and placedaround plate 1. This was repeated for plates 2-7 and the filter. Arectangular ROI was also drawn to assess the background counts. Rawcounts were corrected for background activity and adjusted to counts perminute (cpm).

Aerosol performance is characterized in the table, in terms of the fineparticle fraction (FPF) i.e. % emitted dose with a particle size <5.8μm, mass median aerodynamic diameter (MMAD), geometric standarddeviation (GSD) and the nebulization delivery time.

CEBUD Preparation

The expelled contents of five Pulmicort Respules (1 mg/mL) were combinedtogether. Captisol® (165 mg) on a dried basis was added per Respule usedto the combined contents of the commercial suspension to provide aCaptisol concentration of about 7.5% w/v.

The mixture was vortexed briefly to disperse and dissolve the Captisol®.Then placed on a roller-bed mixer and allowed to mix for two-four orseveral hours. Aliquots of the equilibrated mixture were used to recoverany budesonide retained in the original Respule container, and therecombined together. The mixture was the further equilibrated overnight(˜20 hours) on the roller-bed mixer. After visually checking that allthe suspended solids had dissolved, the required volume of 99mTc-D5PA/saline solution (provided by Medical Physical Department, UHW)was added. So about 1800 of the Radiolabel solution was added to theCaptisol-Enabled® Budesonide Inhalation Solution and vortexed briefly.

Example 23

Determination of the phase solubility curve for corticosteroiddissolution with SAE-CD.

The solubility of coritcosteroid solutions containing SAECD wasdetermined by HPLC chromatography of aliquots from equilibrated filteredor centrifuged corticosteroid solutions as follows.

SAE-CD/steroid solutions were prepared by weighing dry solids of SAE-CD(to provide 0.04 molar) and excess steroid drug (6 mg/mL) together intoa screw-capped vial. A volume of water was aliquoted to each vial(separate vial for each steroid). Intrinsic solubility was determined byweighing excess steroid (6 mg/mL) and adding a volume of water in theabsence of CD. Vials were capped, initially vortexed and sonicated.Vials were then placed on a roller-mixer (model: SRT2; Manufacturer:Stuart Scientific; Serial number: R000100052) or rocker/mixer (Model:LabQuake; Manufacturer: Barnstead/Thermolyne; Serial number:1104010438202). Higher excesses of solid steroid (up to 10 mg/mL) werethen added to any vial where the liquid contents clarified overnight(e.g. prednisolone, hydrocortisone, and prednisone). Samples were rolledand mixed on the roller or rocker for 72 hours. At various times duringthe equilibration, samples were additionally vortexed or sonicatedbriefly (up to 30 minutes). After the designated equilibration time,samples were filtered (0.22 μm, 25 mm, Duropore—PVDF, manufacturer:Millipore) into clean vials except for the intrinsic solubility samplefor Beclomethasone Dipropionate which was centrifuged and thesupernatant transferred to a clean vial. Samples were analyzed byconventional HPLC methods.

Example 24

Preparation of clear liquid formulations containing SAE-CD, budesonideand Salmeterol xinafoate. A formulation of this kind can be preparedaccording to other examples herein containing a combination ofcorticosteroid, SAE-CD and a second therapeutic agent.

Example 25

Preparation of clear liquid formulations containing SAE-CD, budesonideand albuterol sulfate.

A stock solution containing CAPTISOL (40.0 g), water (1 L), citric acid(387.6 mg), sodium citrate (519.6 mg), EDTA (120 mg), and NaCl (6.36 mg)was prepared by mixing the components together.

Budesonide (37.5 mg) and albuterol sulfate (150 mg) were added to analiquot (150 ml) of the stock solution and mixed on a roller mixer atambient temperature until dissolution of all components. The finalsolution was clear, however, it could be filtered if needed. The finalconcentration of components in the formulation was as follows: CAPTISOL(0.018 M); budesonide (5.9 mM or 254 μg/ml); and albuterol sulfate (1mg/ml).

Example 26

Characterization of droplet size distribution of an aerosolized solutionusing a cascade impactor.

The droplet size distribution of an aerosolized solution of theinvention was characterized using an NGI cascade impactor. Approximately0.5 mL of the 1000 μg/mL was placed into a nebulizer, such as theAERONEB GO. The vacuum pump associated with the impactor was turn on.The nebulizer mouthpiece was positioned into the center of the USPinduction port and sealed to it with parafilm. The nebulizer was turnedon until no more vapors were visible. Collection continued for anadditional 45 seconds to ensure complete collection of the sample.Vacuum was turned off and the impactor disassembled. The collection cupsat each stage were extracted with known volumes of HPLC mobile phase andassayed for budesonide. The cumulative amount of budesonide on eachstage was quantified. The percent of drug exiting the nebulizer was 81%,and the percent of drug in the fine particle fraction (<5 μm) was 66%.

Example 27

Determination of total drug output and drug output rate from a nebulizercontaining a liquid of the invention.

A dose collection apparatus was constructed with a 300 ml glass filterunit of the type used to filter HPLC mobile phases. A speciallyfabricated Plexiglas lid covered the reservoir and had an opening toadmit aerosol into the reservoir. The reservoir tapered towards a glassfiber filter supported by a metal mesh. The mesh was contained in aconical glass housing that terminated in a glass tube, which wasattached to a flow controller and vacuum pump. The reservoir and filtersupport housing were clamped together. Two filters were sandwichedtogether in the apparatus to collect the emitted aerosol. This wasnecessary to avoid saturation of the filter, which can result in ittearing with subsequent drug loss into the vacuum, and changes inairflow rate over the course of aerosol collection. The end ofnebulization was judged by intermittent aerosol output and/or a changein pitch of the nebulizer sound. To prevent loss of nebulized budesonideto the atmosphere, air was drawn into the filter faster than it wasejected from the nebulizers. By visually confirming that no aerosolescaped from the back of any nebulizer, a flow rate of 15±5% 1/min wasdeemed satisfactory. This was regulated by a TPK flow controller (CopleyInstruments, 4312) and vacuum pump, and confirmed before each experimentusing a calibrated flow meter.

Example 28

Preparation of a liquid formulation comprising SAE-CD and hudesonide,optionally containing Tween.

A 3 mM citrate buffer at pH 4.5 was added to 2 grams of CAPTISOL and 25mg of budesonide in a serum vial to make the final volume 10 mL. Thesuspension was well mixed by vortexing and sonication. A 20% stocksolution of CAPTISOL without budesonide was also prepared in 3 mMcitrate buffer. These mixtures, along with the buffer were sealed inseparate vials and autoclaved using the 20-minute hold at 121° C. cycle.HPLC analysis of the clear budesonide solution showed the concentrationwas 2100 μg/mL. The 20% CAPTISOL stock solution was used to dilute thesample to 2000 μg/mL. A portion of the above resulting solution wasoptionally diluted with an equal volume of the 3 mM citrate buffer. HPLCanalysis showed the final concentration was 990 μg of budesonide/mL.

The Tween could be added to the above solution as follows. A solution of0.02% Tween was prepared with the autoclaved buffer only solution toform a Tween stock solution for use as a diluent for the abovesolutions. The dilutions for the 10% captisol/1 mg/mL budesonide weredone by weight. Approximately 9 grams of the 20% captsiol/2000 μg/mL wasmixed with ˜9 grams of either the autoclaved buffer only solution or theautoclaved buffer/0.02% Tween solution. These solutions were well-mixed,filtered and reassayed by HPLC.

The hudesonide concentrations of the above formulations were found to be986 μg/mL for the solution without Tween and 962 μg/mL for the solutionwith Tween.

The solutions can be nebulized with any nebulizer; however, with an AERxnebulizer, an initial sample volume of 50 μl can be used. Administrationof this solution with the nebulizer makes it feasible for a therapeuticdose to be administered to a subject in a single puff (a single fullinhalation by a subject) via nebulization.

Example 29

Preparation and dissolution of a lyophilized formulation comprisingSAE-CD and budesonide.

An excess of budesonide, 3.5 mg/mL, was added to 3 L of 30% Captisol in3 mM citrate buffer containing 0.1 mg/mL EDTA. After mixing for 2 days,an additional 1 mg/mL budesonide was added and equilibrated anadditional 4 days. The preparation was filtered through a 0.22μ Duroporefilter and placed in three stainless steel trays in a freeze dryer. Thesolution was frozen at −30° C. for one hour and lyophilized over 30hours to remove essentially all the water. The lyophile was powdered,screened and the powder transferred to a plastic bottle. The finalcomposition contained 8.2 mg budesonide per gram of powder.

When approximately 65 mg of powder was added to 2 mL of water, anessentially clear solution containing the same amount of budesonide asin the reference suspension product was rapidly obtained.

Example 30

Preparation of an aqueous liquid formulation comprising SAE-CD, ethanoland budesonide.

Captisol/Ethanol solutions were prepared by making a stock captisolsolution at 22.2% (˜0.1M) w/v which was diluted with either ethanol orwater in varying amounts to create four solutions of 0, 1, 2, 5% ethanoland about 20% w/v Captisol. Captisol/Ethanol/Budesonide solutions wereprepared by adding dry Budesonide (2.5 mg/mL) to a volume of theprepared Captisol/ethanol solutions and then these were equilibrated ona Labquake for 72 hours. These solutions were filtered (Duropore syringefilters) and analyzed by HPLC to determine the concentration (μg/ml) ofbudesonide dissolved in the formulation.

Example 31

An in-vivo evaluation of a dosage form according to the invention wasconducted by performing a pharmacokinetic analysis on budesonide afteradministration of the dosage form by nebulization.

A four dog crossover study to compare pulmonary delivery of budesonidevia Pulmicort Respules®, and a Captisol-Enabled® budesonide formulationusing a Pari LC air-jet nebulizer was conducted. The purpose of thestudy was to determine, by HPLC-MS/MS analysis, the pharmacokinetics ofbudesonide the plasma, following nebulization of a budesonide suspension(Pulmicort Respules®, AstraZeneca, reference formulation) and aCaptisol-Enabled® budesonide inhalation solution (test formulation) inhealthy dogs. Dosing was conducted using a Pari LC Star air-jetnebulizer and anesthesia masks.

The study dosage forms consisted of: 1) 1 mg Budesonide as two 2 mL×0.25mg/mL Pulmicort Respules®; or 2) 1 mg Budesonide as two 2 mL×0.25 mg/mLPulmicort Respules to which 5% w/v Captisol® was added.

Prior to dosing, two capsules containing activated charcoal followedwith approximately 30 mL of water were administered to each dog to blockoral absorption of the budesonide. Each dog received each of the studyadministrations one week apart in a randomized manner. Administrationtime was the same for each formulation (23.9 min. for test formulationand 23.4 min. for reference). Blood was collected periodically up to 8hours, the plasma was isolated and stored at −70° C. until analysis.After each administration, the nebulizer and test solution containerwere assayed for residual budesonide to determine the amount of drugdelivered.

Example 32

To investigate how the incorporation of Captisol at 5% w/v intoPulmicort Respules impact performance of different types of nebulizers.

The emitted dose of budesonide from four different nebulizers (PARI LCPLUS (air jet), OMRON MICROAIR NE-U22, AIRSEP MYSTIQUE (ultrasonic),AEROGEN AERONEB) was determined. The package insert-approved Pari airjet system was used as the benchmark to judge performance of the othernebulizers. The emitted dose was from 1.25 to 3.7 times higher whenCaptisol was added to the budesonide suspension. The Emitted dose (ED)was determined by:

-   -   1) Drawing the nebulized formulations through a 300 mL glass        filter apparatus at 15 l/min, and collecting drug on double or        triple layers of glass fiber depth filter and the interior        walls. Collection was stopped every two minutes, the budesonide        quantitatively recovered, and filters were changed to prevent        filter saturation or alterations in airflow. Budesonide recovery        was quantified by HPLC; and/or    -   2) Summing the amount of budesonide on the cascade impactor        stages after nebulization.

The results are detailed below. (ND means “not determined.)

Total Delivered (ED) Total Delivered (ED) (μg, mean & SD), (μg, mean &SD), Formulation Filter¹ Impactor² Pari LC Plus (Air Jet) Listed in thePulmicort package insert Pulmicort 171.5 ± 6.3  137.8 ± 14.9 Pulmicort +247.4 ± 11.3 172.4 ± 6.6  5% Captisol Omron MicroAir NE-U22 Pulmicort179.9 ± 17.2 168.8 ± 30.1 Pulmicort + 380.1 ± 8.5  349.6 ± 10.0 5%Captisol AirSep Mystique (Ultrasonic) Pulmicort 32.9 ± 6.4 ND*Pulmicort + 120.8 ± 19.6 ND* 5% Captisol Aerogen AeroNeb Pulmicort 90.7± 4.5 ND* Pulmicort + 301.2 ± 19.5 ND* 5% Captisol

All documents cited herein are each incorporated by reference herein inits entirety. The above is a detailed description of particularembodiments of the invention. It will be appreciated that, althoughspecific embodiments of the invention have been described herein forpurposes of illustration, various modifications may be made withoutdeparting from the spirit and scope of the invention. Accordingly, theinvention is not limited except as by the appended claims. All of theembodiments disclosed and claimed herein can be made and executedwithout undue experimentation in light of the present disclosure.

1. A method of treating a disease or disorder of the airways in asubject in need thereof, comprising administering via inhalation to thesubject an aqueous liquid formulation comprising an aqueous liquidcarrier, a sulfoalkyl ether cyclodextrin (SAE-CD), and a dose ofcorticosteroid dissolved therein, wherein the formulation provides anenhanced pharmacokinetic profile over a suspension based formulationadministered under similar conditions.
 2. A system comprising: anebulizer equipped with a reservoir; and an aqueous liquid formulationcomprising an aqueous liquid carrier, solubility enhancer, and a dose ofcorticosteroid, wherein, during nebulization of the formulation, thenebulizer provides a percentage decrease in the rate of increasingconcentration of corticosteroid in the formulation in the reservoir ascompared to the rate of increasing concentration of a suspension-basedformulation nebulized with the nebulizer under similar conditions. 3.The method of claim 1, wherein the dose of corticosteroid is present inan amount sufficient to provide a mean plasma AUC_(t) of 160-1600pg*h/ml, and wherein the molar ratio of SAE-CD to corticosteroid isgreater than 10:1.
 4. The method of claim 1, wherein the formulationcomprises 25-400 μg of a corticosteroid, and wherein the formulationprovides a plasma AUC_(t) of 150-1600 pg*h/ml for the corticosteroid. 5.The method of claim 1, wherein the formulation comprises at least 25 μgof corticosteroid, and wherein the formulation provides a plasmaAUC_(t), normalized for dose of corticosteroid, of at least 6(pg*h/ml)/μg of corticosteroid.
 6. The method of claim 1, wherein theformulation comprises at least 25 μg of corticosteroid, and wherein theformulation provides a plasma AUC_(i), normalized for dose ofcorticosteroid, of at least 8 (pg*h/ml)/μg of corticosteroid.
 7. Themethod of claim 1, wherein the SAE-CD is a compound of the formula I:

wherein: n is 4, 5 or 6; R₁, R₂, R₃, R₄, R₅, R₆, R₇, R₈ and R₉ are each,independently, —O— or a —O—(C₂-C₆ alkylene)-SO₃ ⁻ group, wherein atleast one of R₁-R₉ is independently a —O—(C₂-C₆ alkylene)-SO₃ ⁻ group, a—O—(CH₂)_(m)SO₃ ⁻ group wherein m is 2 to 6, —OCH₂CH₂CH₂SO₃ ⁻, or—OCH₂CH₂CH₂CH₂SO₃ ⁻); and S₁, S₂, S₃, S₄, S₅, S₆, S₇, S₈ and S₉ areeach, independently, a pharmaceutically acceptable cation.
 8. The methodof claim 1, wherein the SAE-CD is a compound of the Formula II(SAEx-α-CD), wherein “x” ranges from 1 to 18; of the Formula III(SAEy-β-CD), wherein “y” ranges from 1 to 21; or of the Formula IV(SAEz-γ-CD), wherein “z” ranges from 1 to 24, and wherein “SAE”represents a sulfoalkyl ether substituent, and the values “x”, “y” and“z” represent the average degree of substitution in terms of the numberof sulfoalkyl ether groups per CD molecule.
 9. The method of claim 1,wherein the Cmax provided by the corticosteroid formulation is at least1.5 fold higher than the Cmax provided by the suspension-basedformulation when the dose of corticosteroid in the formulation is about2 fold lower than the dose in the suspension.
 10. The method of claim 1,wherein the formulation provides a C_(max) (pg/ml) of 1600 to 1800 on adose non-normalized basis.
 11. The method of claim 1, wherein, on thebasis of the normalized nominal dose of corticosteroid, the C_(max)provided by the corticosteroid formulation is at least 1.7 fold higherthan the C_(max) provided by the suspension-based formulation when thedose of corticosteroid in the formulation and the suspension isapproximately the same.
 12. The method of claim 1, wherein theformulation provides an AUC_(inf) (pg*h/ml) of 250 to 2500 on a dosenon-normalized basis.
 13. The method of claim 1, wherein, on the basisof the non-normalized dose of corticosteroid, the AUC_(inf) provided bythe corticosteroid formulation is 1.5 to 3.5 fold higher than theAUC_(inf) provided by the suspension-based formulation when the dose ofcorticosteroid in the formulation and the suspension is approximatelythe same.
 14. The method of claim 1, wherein the AUC_(inf) provided bythe corticosteroid formulation is at least 1.5 fold higher than theAUC_(inf) provided by the suspension-based formulation when the dose ofcorticosteroid in the formulation is about 2 fold lower than the dose inthe suspension.
 15. The method of claim 1, wherein the formulationprovides an AUC_(0-8 hr) (pg*h/ml) of 2000 to 3000 on a dosenon-normalized basis for the corticosteroid.
 16. The method of claim 1,wherein the corticosteroid is budesonide.
 17. The method of claim 1,wherein the corticosteroid is selected from the group consisting ofbeclomethasone dipropionate, beclomethasone monopropionate, budesonide,ciclesonide, desisobutyryl-ciclesonide, flunisolide, fluticasonepropionate, fluticasone furoate, mometasone furoate, icomethasoneenbutate, tixocortol 21-pivalate, and triamcinolone acetonide.
 18. Themethod of claim 1, wherein the formulation is administered with anebulizer selected from the group consisting of an air jet nebulizer,ultrasonic nebulizer, electronic nebulizer, vibrating membranenebulizer, vibrating mesh nebulizer, vibrating plate nebulizer, anebulizer comprising a vibration generator and an aqueous chamber, and anebulizer comprising a nozzle array.
 19. A method of preparing acorticosteroid formulation, comprising high shear mixing acorticosteroid, SAE-CD, and an aqueous liquid carrier, wherein theSAE-CD is present in an amount sufficient to dissolve thecorticosteroid.
 20. The method of claim 19, wherein high shear mixing isconducted with a sonicator, narrow gauge syringe(s), mixer/homogenizer,rotor-stator mixer, or saw tooth mixer.